See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/283294232

The Endoplasmic Reticulum

Chapter · December 2016

DOI: 10.1016/B978-0-12-394447-4.20012-6

CITATIONS READS

3 27,985

2 authors:

Eelco van Anken Roberto Sitia

San Raffaele Scientific Institute Università Vita-Salute San Raffaele
89 PUBLICATIONS   2,548 CITATIONS    265 PUBLICATIONS   15,139 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Cloning od new genes related to endoplasmic reticulum oxidoreductin View project

A novel Correlative Light and Electron Microscopy approach to investigate dynamic complexity of ER-to-Golgi transport View project

All content following this page was uploaded by Eelco van Anken on 25 September 2017.

The user has requested enhancement of the downloaded file.

 Author's personal copy

ORGANELLES: STRUCTURE AND FUNCTION

Contents
The Endoplasmic Reticulum
Intermediate Compartment: A Sorting Station between the Endoplasmic Reticulum and the Golgi Apparatus
Golgi and TGN
Early Endosomal Compartments
The Late Endosome
Signaling from Endosomes
Conventional and Secretory Lysosomes
Lysosome-Related Organelles
At the Center of Autophagy: Autophagosomes
Peroxisomes
Lipid Droplets
Mechanisms and Functions of Mitochondrial Dynamics
Synaptosomes and Synaptic Vesicles
Extracellular Vesicles
Nuclear Organization (Nuclear Structure and Dynamics)
Nuclear Pores
Plastids: The Anabolic Factories of Plant Cells

The Endoplasmic Reticulum

E van Anken and R Sitia, Vita Salute San Raffele University, Milan, Italy
r 2016 Elsevier Inc. All rights reserved.

Introduction entered the canon of cell biology as a distinct organelle sep-

Robert Hooke discovered that tissues consist of cells already in
1665. Fellow microscopists at that time, Van Leeuwenhoek
and Bauer, realized that smaller structures were discernable
within the cell, in particular the vacuole and the nucleus. Yet,
the terminology for these cell compartments stems from the
nineteenth century, when several other subcellular structures
were discovered. Möbius then coined the name ‘organula’ – a
term that was later simplified to ‘organelles’ – to describe these
subcellular compartments. Just before the turn of the century,
Camillo Golgi identified the organelle that still bears
his name.
At the very beginning of the twentieth century, a student of
Golgi, Emilio Veratti, found yet another subcellular structure
in muscle cells, which he claimed to be distinct from the Golgi
apparatus (Veratti, 1961). Veratti’s work failed to convince his
contemporaries, however. About 50 years later, the advent of
the electron microscope finally made the unambiguous iden-
tification of Veratti’s elusive organelle possible when Keith
Porter and colleagues observed a network (reticulum) inside
the cytoplasm of eukaryotic cells (endoplasmic). He teamed
up with George Palade to obtain high-resolution images
(Figure 1) and in 1954 the endoplasmic reticulum (ER)
arated from the cytosol by virtue of a membrane and forming
a complex tubular or sheet-like network (Palade and Porter,
1954).
The Palade lab then developed a method – which still is
widely used today – to obtain ER-enriched cell fractions, called
microsomes (Figure 1). The physical sheering of cells causes
ER vesiculation and ER-derived vesicles are then purified by
differential centrifugation (Palade and Siekevitz, 1956). The
combination of microscopy and biochemical assays exploiting
microsomes initiated a cascade of discoveries on the function
and morphology of the ER and its specialized derivative in
muscle cells: the sarcoplasmic reticulum (SR). In the early
1960s, Palade and coworkers proved by pulse chase analysis
using radiolabeling that the ER is the first station along the
secretory pathway (Siekevitz and Palade, 1960). Günther
Blobel (a pupil of Palade) and colleagues elucidated in the
1970s that client proteins enter the ER by virtue of a signal
peptide (Blobel and Dobberstein, 1975).
Another milestone in ER research came when genetic
screens in yeast, pioneered by Randy Schekman and colleagues
(Novick et al., 1980), initiated the characterization of the
molecular machinery of the secretory pathway. A long list of
the so-called sec proteins was identified, including several that

156 Encyclopedia of Cell Biology, Volume 2 doi:10.1016/B978-0-12-394447-4.20012-6

 Author's personal copy
Organelles: Structure and Function: The Endoplasmic Reticulum
(a) (b)
Figure 1 (a) One of the first sightings of the ER by electron microscopy at high resolution (n ¼nucleus, e¼ER). Reproduced from Palade, G.E.,
Porter, K.R., 1954. Studies on the endoplasmic reticulum. I. Its identification in cells in situ. Journal of Experimental Medicine 100, 641–656.
(b) An original slide produced in the 1960s in the Palade laboratory of an electron micrograph of microsomes, personal collection of Professor
Jacopo Meldolesi, a former collaborator of Palade. (c) Magnification of the area around the handwritten size bar of the slide shown in the middle
panel; note that the microsomes evidently are ribosome-studded.
are fundamental to ER function. Through the work of Ari
Helenius and coworkers it became clear in the late 1980s/early
1990s that stringent quality control is exerted on protein
folding in the ER (Hurtley et al., 1989). At around the same
time, pioneering work from Kazutoshi Mori and Peter Walter
(a pupil of Blobel) initiated research on the unfolded protein
response (UPR) (Cox et al., 1993; Mori et al., 1993), which is
key for homeostasis of the ER. In this article we summarize the
current understanding of this fascinating organelle.
Origin and Function of the ER
The hallmark of eukaryotic cells is the packing of DNA into the
nucleus, a compartment that is bounded by the nuclear en-
velope. The nuclear envelope is continuous with the ER, which
suggests that the ER was present in eukaryotic cells right from
their ‘inception.’
As the first station along the exocytic pathway, the ER serves
as an anticipation of the extracellular world, to prepare mol-
ecules to exert their function in a different environment. As such,
it is functionally related to the periplasm, the space enclosed
between the inner membrane and cell wall of gram-negative
bacteria. In analogy, portions of the ER (the so-called cortical
ER) are positioned directly below the cell surface also in eu-
karyotic cells. Both the periplasm and the ER serve to groom
proteins that are destined to be part of the cell membranes or to
be secreted. Instead of being translocated directly across the
plasma membrane, these proteins enter the ER or periplasm
first. Here, under the supervision of resident chaperones and
enzymes, they attain their native structure before being deployed
to the extracellular space. Altogether, these mechanisms warrant
efficient production and transport, while simultaneously en-
suring the fidelity of the protein secretome, which comprises a
third of the eukaryotic proteome, equaling in humans approxi-
mately 18 000 gene products (Chen et al., 2005).
Ribosomes that are translating proteins destined for the
secretory pathway dock onto the cytosolic face of the ER
membrane. These ribosomes give a ‘rough’ appearance to the
ER, typical of protein secreting cells. Accordingly, the ribo-
some-studded parts of the ER therefore are referred to as
‘rough ER,’ while the portion devoid of ribosomes as ‘smooth
ER.’ The majority of ‘ER client’ proteins enter co-translationally
into the organelle. Some proteins can enter after their synthesis
157
(c)
is completed (posttranslationally). In either case, the proteins
traverse the ER membrane through the so-called translocon
complex. The latter’s main component, Sec61, has an evo-
lutionary equivalent in gram-negative bacteria (Keenan et al.,
2001) that allows proteins to reach the periplasm, which
supports the notion that the ER has a periplasmic-like origin.
Next to being the cradle of secretory and cell surface pro-
teins, the ER is the most important site of (membrane) lipid
synthesis in the eukaryotic cell (Fagone and Jackowski, 2009).
Accordingly, a vast repertoire of enzymes devoted to lipid syn-
thesis resides in the ER. Other organelles depend on membrane
lipid supply from the ER, either through vesicular transport or
through lipid transfer. The numerous contact points between ER
and other organelles likely are sites of such lipid transfer, even
though the membranes of the ER and the acceptor organelles
are juxtaposed and not continuous. In other organelles the
membrane lipids that are derived from the ER often are further
derivatized or asymmetrically distributed between membrane
leaflets according to need (Vance et al., 1977).
In most eukaryotic cells, the ER also is the major store
of intracellular Ca2 þ ions and most ER resident proteins in
fact are low-affinity/high-capacity calcium-binding proteins
(Meldolesi and Pozzan, 1998). A wealth of signaling pathways
exploits the Ca2 þ gradient across the ER membrane (Berridge,
2002). Typically, signaling cascades drive the opening of Ca2 þ
channels to sustain a burst of Ca2 þ through vicinal mito-
chondria and in the cytosol, which, in turn, serves to propagate
the signal. In most cases, there is a rapid reuptake of Ca2 þ
back into the ER, ensuring that Ca2 þ signaling can start anew.
The most sophisticated Ca2 þ -driven signaling cycles occur at
the sarcoplasmic reticulum to sustain muscle contraction
(Endo, 1977).
Morphology of the ER
The ER is the most extended organelle in many eukaryotic
cells: on average it occupies 10% of the cellular volume, but
this figure is larger in plasma cells, exocrine pancreas, and
other professional secretory cells. The tubules and sheet-like
structures (both rough and smooth) form a continuous
membrane system that includes the nuclear envelope (Voeltz
and Prinz, 2007). Tubules can be highly curved and branched
to form a polygonal network extending throughout the whole
 Author's personal copy
158 Organelles: Structure and Function: The Endoplasmic Reticulum

cytosol. ER sheets can be prominent around the nucleus, and
may become predominant in some cell types dependent on
their function (Figure 2).
Alterations in membrane curvature form the basis of
the different morphologies of the ER. For instance, ER tubules
are cylindrical with a diameter of 30–100 nm, while sheets
are virtually flat. The width of the ER lumen in sheets is
restricted to approximately 50–100 nm (Voeltz and Prinz,
2007), but occasionally sac-like structures may develop,
bulging from the ER sheet and tubular network, in particular
when cells sustain bulk protein secretion or when aberrant
proteins aggregate into ER resident deposits (Kopito and Sitia,
2000).
In recent years insights into the mechanisms that are re-
sponsible for ER morphology have greatly improved. A num-
ber of proteins have been identified that determine its shape.
Wedge-like proteins, reticulons and other similar proteins,
insert into the ER membrane and dictate the typical curvature
found in ER tubules and at the edges of ER sheets, including
the nuclear envelope at the sites of nuclear pores (Voeltz et al.,
2006). Proteins driving formation of sheet-like structures
include CLIMP-63, a transmembrane protein whose ER lu-
minal domains can homodimerize with another CLIMP-63
molecule protruding into the lumen from the opposing
membrane in the ER sheet, and so determining the typical
width (Klopfenstein et al., 2001). Other proteins, for example,
kinectin and p180, with bulky rod-like structures facing the
cytosol seem to be required for stabilizing the flatness of sheets
(Lin et al., 2012; Figure 3).
While strictly organized in rough and smooth, tubular, and
sheet-like, the ER is highly dynamic at the same time. The
network is constantly reshaped through fission and fusion
events, in which GTPases of the atlastin and Rab families are
involved (Hu et al., 2009; English and Voeltz, 2013). Re-
shaping of the ER occurs hand in hand with movements of ER
elements along the cytoskeleton that are facilitated by motor
proteins (Goyal and Blackstone, 2013).
Depending on the cell type and functional requirements,
the shape, organization, and size of the ER varies considerably.
In muscle cells, for instance, the sarcoplasmic reticulum is a
highly organized form of smooth ER devoted to synchronize
myofibril contraction (Endo, 1977), while the rough portion

Nuclear
pore
Nuclear
envelope
Nucleus
Cisternae

Rough Cisternal
endoplasmic space
reticulum
Ribosomes
Smooth
endoplasmic Peripheral
reticulum sheet

Tubules

Figure 2 Schematic representation of the ER in animal cells. Reproduced from Goyal, U., Blackstone, C., 2013. Untangling the web: Mechanisms
underlying ER network formation. Biochimica et Biophysica Acta 1833, 2492–2498.

Ribosomes

Scaffolding
Atlastins protein

Smooth ER sheet
ER tubule lumen
ER
lumen Reticulons
Reticulons Luminal spacer
(CLIMP-63)

Figure 3 Wedge-shaped proteins such as reticulons facilitate membrane curvature in ER tubules or at edges of sheets. Spacers such as
CLIMP-63 maintain thickness of sheets. Reproduced from Goyal, U., Blackstone, C., 2013. Untangling the web: Mechanisms underlying
ER network formation. Biochimica et Biophysica Acta 1833, 2492–2498.
 Author's personal copy
Organelles: Structure and Function: The Endoplasmic Reticulum 159

of the ER is marginal. In liver cells, the smooth ER is also
extensively developed and a major site of detoxifying and
synthetic enzymes. In professional secretory cells such as
thyrocytes that produce thyroglobulin, acinar cells, which
produce digestive enzymes, etc. the rough ER instead is pre-
eminent. Antibody producing plasma cells are prototypes of
cells with an expanded rough ER (Figure 4).
Membrane Contact Sites of the ER with Other
Organelles
Subdomains of the ER make close contact with virtually all
other cellular structures, foremost with the nucleus, as the
nuclear envelope is continuous with the ER. Contacts of ER
Figure 4 Electron micrograph of a plasma cell, which has an
expanded ER to sustain antibody production and secretion. Courtesy
of the late Dr. Carlo E. Grossi.
with other organelles are maintained by various tethering
complexes that form a bridge between the apposing mem-
branes. ER–plasma membrane contacts are predominant in
yeast in which cortical ER represents up to 50% of the or-
ganelle. In metazoans ER–plasma membrane contacts are less
abundant, but the interaction between STIM in the ER mem-
brane and Orai in the plasma membrane is of particular
interest. Together, STIM and Orai mediate store-operated
Ca2 þ entry from the extracellular space across both mem-
branes into the ER lumen (Stefan et al., 2013; Figure 5).
Likewise, protein-mediated tethering of ER to mito-
chondria sustains close contacts, which are referred to as
mitochondria-associated membranes (MAM). In metazoans,
these ER–mitochondria contact sites are key for Ca2 þ transfer
and, thus, for signaling: the opening of IP3 receptor channels
at the MAM leads to Ca2 þ efflux, which is readily imported
into the mitochondrial matrix through porins and mitochon-
drial calcium uniporters (MCUs) to initiate signaling. Excess
Ca2 þ is pumped back from the cytosol into the ER by SERCA
(sarco-endoplasmic reticulum Ca2+ ATPase) or across the PM
by PMCA (plasma membrane Ca2+ ATPase) (Figure 5). Ca2 þ
signaling activates some enzymes of the Krebs cycle and, thus,
is key for overall metabolism. Yet, unrestrained Ca2 þ transfer
to mitochondria initiates apoptosis (Naon and Scorrano,
2014).
While Ca2 þ shuttling through ER–mitochondria contacts
seems less important in yeast, the contact sites are key for
lipid transfer in all eukaryotic cells (Helle et al., 2013). Mito-
chondria do not synthesize membrane lipids and there is no
vesicular transport or any other membrane continuity between
mitochondria and the endomembrane system. Thus, mito-
chondria must be furnished with membrane lipids through
lipid transfer proteins. Yet, so far it remains unclear how the
lipid transfer process at the MAM is organized.
The ER establishes tether-mediated contacts also with the
Golgi, lysosome, and endosomes. Their main function seems

Extracellular space

PMCA pump
ER SERCA pump
Orai channels

STIM

IP3R
VDAC/Porin
MCU
Mitochondrion
Tether, for example,
mitofusin

Figure 5 Organization of Ca2 þ signaling through ER–PM and ER–mitochondria contact sites. Blue represents the local [Ca2 þ ]. A Ca2 þ gradient
is established by SERCA (red) and PMCA (yellow) pumps that pump Ca2 þ ions from the cytosol into the ER or, respectively, into the extracellular
space. ER–PM contacts facilitate replenishing of Ca2 þ stores in the ER from the extracellular space. STIM (dark green) tethers the ER membrane
to the PM at sites where Orai channels are, such that little Ca2 þ leaks to the cytosol proper. Ca2 þ is released from the ER through IP3R channels
(tan) mostly at sites, where tethers such as mitofusin (orange) maintain close ER–mitochondria contacts. Ca2 þ is readily taken up by mitochondria
via VDAC (Voltage-dependent anion channels)/Porin (blue) and MCU (light green) channels. Adapted from Helle, S.C., Kanfer, G., Kolar, K., et al.,
2013. Organization and function of membrane contact sites. Biochimica et Biophysica Acta 1833, 2526–2541.
 Author's personal copy
160 Organelles: Structure and Function: The Endoplasmic Reticulum

Signal-recognition particle Ribosome

SRP-bound to signal
peptide

ER membrane
Signal peptide
Accessory proteins
of the translocon
complex
Protein Translocating
SRP-receptor channel polypeptide chain
(‘translocon’)
Figure 6 Proteins destined for the secretory pathway display an N-terminal signal peptide, which is recognized by SRP. The ternary complex of
the ribosome, the nascent chain, and SRP is recognized by the SRP receptor and docks onto the translocon, whereupon co-translational
translocation into the ER begins.

Endoplasmic reticulum
lumen
Single-spanning Multi-spanning
Soluble membrane proteins GPI-anchored membrane proteins
Type I Type II Tail- Type I Type II
anchored

C C
N N C N N
C

N N
C N
C

Figure 7 Topology of ER client proteins. From many ER client proteins the signal peptide (light blue) is removed by signal peptidase (scissors).
If the protein has no other hydrophobic stretch, it will be a soluble protein. If the protein has one or more hydrophobic stretches (green) it will be
membrane embedded. Tail-anchored proteins have negligible ectodomains at the C-terminus. Some proteins obtain a GPI-anchor, consisting of
lipid moieties (wiggled lines) of phosphoinositol (yellow hexagon) that allow membrane embedding coupled via glucosamine (green square), three
mannose moieties (pink) and phosphoethanolamine to the polypeptide chain.

to be the transfer from the ER of specific lipids that are in
shortage in the target membranes (Helle et al., 2013). Overall
storage of lipids takes place in lipid droplets, and it comes as
no surprise that lipid droplets are intimately linked with the
ER as well (Xu et al., 2012). The ER moreover is the source of
membrane lipids for the peroxisome. While de novo peroxi-
some formation initiates with pre-peroxisomal structures de-
rived from the ER (van der Zand et al., 2012), it is unclear
whether mature peroxisomes require lipid transfer from the ER
through contact sites. Finally, it is still highly debated what is
the source of autophagic membranes, which also may turn out
to originate either directly or indirectly from the ER.
Translocation and Membrane Embedding of ER Client
Proteins
Most client proteins enter the ER co-translationally by virtue of
a hydrophobic signal peptide generally located at their
N-termini (Blobel and Dobberstein, 1975). As such, the signal
peptide emerges first from the ribosome during translation,
whereupon it binds to the signal recognition particle (SRP)
(Walter et al., 1981). SRP binding temporary arrests elongation
of the nascent polypeptide and guides the translating ribo-
some to the SRP receptor, which is associated with the trans-
locon complex (Gilmore et al., 1982). The net result is that
translation is now coupled to translocation (Figure 6). The
hydrophobic signal peptide is laterally released from the
translocon into the ER membrane, while the remainder of
the ER client nascent chain reels into the ER lumen with the
assistance of resident chaperones (Tyson and Stirling, 2000).
Often the signal peptide is then removed by signal peptidase
(Blobel and Dobberstein, 1975), but when it is not cleaved off,
the signal peptide serves as a membrane anchor (Figure 7).
Translocation continues unless the translocon encounters
another hydrophobic stretch in the nascent chain. In that case,
again, elongation slows down as the newly made hydrophobic
segment is laterally released into the ER membrane to serve as
 Author's personal copy
Organelles: Structure and Function: The Endoplasmic Reticulum 161

a transmembrane domain. The remainder of the nascent chain

will then be cytosolic unless the translocon encounters yet
again a hydrophobic stretch, in which case this segment also Endoplasmic reticulum
lumen
will be embedded into the ER membrane. The next portion of
the nascent ER client will then enter the ER lumen again, etc.
As such, multiple membrane spanning proteins are ‘stitched’
Reduced
into the ER membrane and the topology is determined of
PDI SH
cytosolic domains, transmembrane domains, and the so-called
SH
ectodomains inside the ER lumen. Depending on the ‘topo- S S
genesis’ membrane spanning proteins can be oriented with
their N-terminus inside or outside the ER lumen. The first are S S

Isomerization
referred to as type I membrane proteins and the latter as type II
(Goder and Spiess, 2001; Figure 7).
A few examples are known of small proteins that traverse Reduced
the translocon when they are fully synthesized, including the PDI SH S
honeybee venom precursor (Zimmermann and Mollay, 1986). SH S
To sustain such posttranslational translocation, dedicated ac-
S
cessory proteins of the translocon complex, including Sec63,
S
are required (Lang et al., 2012). Also tail-anchored proteins are
posttranslationally inserted into the ER membrane, because

Oxidation
the hydrophobic domains that will serve as membrane an-
chors are at the C-terminus, and thus emerge from the ribo- Oxidized SH
some only once these proteins are already fully synthesized PDI S
(Figure 7). For tail-anchored protein insertion into the ER S SH
membrane TRC-40 (Get3 in yeast) is key to shuttle them from
the cytosol to a receptor complex/insertion machinery in the SH
ER membrane, consisting of WRB and CAML (Get1/Get2 in
yeast) (Schuldiner et al., 2008; Wang et al., 2014). SH

Posttranslational Modifications of ER Client Proteins
Besides proteolytic cleavage of the signal peptide, three other
common posttranslational modifications of client proteins (or
rather their ectodomains) are special to the ER: N-linked gly-
cosylation, insertion of disulfide bonds, and addition of gly-
colipid tail anchors. Client proteins may obtain in the ER a
glycosylphosphatidylinostil (GPI) anchor covalently attached
to their C-termini (Udenfriend and Kodukula, 1995). This
glycolipid moiety is another means to ensure that the client
protein remains associated with the membrane (Figure 7).
Like in the bacterial periplasm, free cysteine residues of
client proteins are oxidized in the ER such that two cysteine
residues are covalently linked to each other (Figure 8). Di-
sulfide bonds can be formed within (intrachain disulfides) or
between (interchain disulfides) client proteins. The latter are
important in stabilizing multimeric proteins. Tightly regulated
disulfide interchange reactions, driven by protein relays,
regulate oxidative protein folding, a process intimately linked
to redox signaling, in that reactive oxygen species can be
generated as byproducts (Tu and Weissman, 2004).
ER client proteins moreover can be glycosylated when an
asparagine (N) residue is followed by any residue but proline
(X) and then a serine or threonine (NXS/T). The glycan, con-
sisting of three glucoses, nine mannoses, and two N-acetyl
glucosamines (Figure 9), is transferred en bloc from dolichol
precursors. Glycosylation occurs most often co-translationally,
as soon as a site emerges in the ER lumen and is recognized by
oligosaccharyl transferase (Helenius and Aebi, 2004). Not all
potential acceptor sites are always used, however. In the prion
Figure 8 ER client proteins obtain disulfide bonds. PDI or PDI-like
proteins catalyze the reaction, but also rearrange aberrant disulfide
bonds.
protein, for instance, glycosylation competes with disulfide
bonding and GPI addition, which increases the number of
isoforms that can be produced (Orsi and Sitia, 2007). Soon
after their en bloc addition to proteins in the ER, N-linked
glycans are sequentially trimmed by resident glucosidases and
mannosidases (Ellgaard and Helenius, 2003), providing im-
portant cues for quality control (see below).
Several other posttranslational modifications can occur in
the ER. Some ER client proteins obtain mannose moieties
that are covalently linked to serine or threonine residues. Such
O-mannosylation occurs predominantly in serine/threonine-
rich domains of ER client proteins (Praissman and Wells,
2014). Relevant to human pathology is hydroxylation of
proline and lysine residues in collagens. The responsible ER
resident hydroxylases use vitamin C as an obligatory cofactor,
which nicely explains the pathogenesis of scurvy.
Protein Folding in the ER
Newly synthesized proteins progressively adopt conformations
that are energetically more favorable, until the correctly folded
‘native’ conformation is reached. Anfinsen’s axiom that folding
information is contained in the protein sequence undoubtedly
holds true (Anfinsen, 1973). Yet, metastable nonnative
 Author's personal copy
162 Organelles: Structure and Function: The Endoplasmic Reticulum

Endoplasmic reticulum
lumen
N-glycan addition Unsuccessful fold
G
N M To proteasome

G
Gluc I + II
M
CNX

G
Gluc II Man I
M M

Successful fold
UGGT
N N To ER exit sites
G

Figure 9 ER client proteins obtain N-glycans, consisting of 2 GlucNAc moieties (green squares), 9 mannose moieties (pink circles), and 3
glucose moieties (blue diamonds). Glucoses are trimmed by glucosidases I and II and monoglucosylated glycans allow association with lectin
chaperones – calnexin (CNX) is shown here – Gluc II then removes the last glucose and the ER client will either go to ER exit sites (if successfully
folded) or it will re-associate with CNX, because UGGT re-added a glucose. Repetitive (de-)glucosylation cycles may occur until mannose trimming
leads to association with lectins like EDEM that target the client for ERAD by the proteasome. Adapted from Mitchell, W.B., Li, J., French, D.L.,
Coller, B.S., 2006. αIIbβ3 biogenesis is controlled by engagement of αIIb in the calnexin cycle via the N15-linked glycan. Blood 107, 2713–2719.

conformations may be adopted, often hindering productive
completion of the folding process. This phenomenon is par-
ticularly evident when folding goes hand in hand with di-
sulfide bond formation. Not surprisingly, therefore, Anfinsen
identified protein disulfide isomerase (PDI) as a crucial cata-
lyst through his pioneering work on RNase folding (Gold-
enberger et al., 1963). PDI can reshuffle disulfide bonds such
that client proteins efficiently attain their native structure
(Figure 8). The folding of ER client proteins often is particu-
larly complex and, hence, the ER hosts an impressive repertoire
of molecular chaperones and foldases.
Classical molecular chaperones in the ER are functionally
similar to counterparts in other folding environments such as
the HSP70 family member BiP (Kar2 in yeast). Similar to other
HSP70s in the cytosol, mitochondria, etc., BiP transiently
binds hydrophobic patches of ER client proteins and thereby
protects them from undergoing inappropriate interactions
with other proteins (Mayer and Bukau, 2005). BiP is an
ATPase and cycles of client binding and release require ATP
hydrolysis. Several J-domain containing co-chaperones
(ERdj1–5), but also nucleotide exchange factors, modulate
BiP’s activity and help determine its clientele (Shen et al.,
2002; Steel et al., 2004). A distant HSP70-like relative in the
metazoan ER is GRP170 that often acts in tandem with BiP,
most notably assisting translocation of client proteins (Steel
et al., 2004). Similarly, GRP94, the HSP90 equivalent in the
metazoan ER, guides maturation of a limited repertoire of
client proteins likely with assistance from co-chaperones of the
CNPY family (Eletto et al., 2010; Liu et al., 2010). Surprisingly
perhaps, the ER lacks members of small HSP (although they
are present in plant ER) and the HSP60 families of chaperones.
Foldases are enzymes that catalyze folding steps. For the
folding of many client proteins cis/trans-isomerization of
proline residues is the rate-limiting step. To smoothen the
process of prolyl isomerization, the ER harbors a number of
peptidylprolyl isomerases (PPIases), similar to other folding
compartments. The most prominent PPIase in the ER is
cyclophilin B, but also a few members of the FKBP family are
present (Galat, 2003).
Unique to the ER are foldases of the PDI family that cata-
lyze formation of disulfide bonds (Figure 8). Dependent on
the cell type, various PDI family members are present, each
likely with (slightly) distinct oxidative properties and clientele.
Notable PDI family members include ERp57, ERp72, and P5
(Appenzeller-Herzog and Ellgaard, 2008). As in the case of
RNAse, PDIs can reshuffle disulfide bonds, exchanging ab-
errant for native disulfide bonds. The de novo disulfide bond
formation requires transfer of disulfide bonds from PDIs to
client proteins. This process entails that PDIs are constantly
reoxidized. In yeast, reoxidation of PDI is the exclusive task of
Ero1, which uses O2 as terminal electron acceptor, thereby
generating H2O2 (Tu and Weissman, 2004). In metazoans,
ERO1 proteins share the burden of generating reoxidizing
equivalents to sustain disulfide bond formation with peroxir-
edoxin IV, glutathione peroxidases 7 and 8, and vitamin K
epoxide reductase (Bulleid and Ellgaard, 2011).
The glycans play a unique role in the folding process of ER
client proteins. Once the first two glucose moieties are removed
by glucosidases, lectin chaperones of the calnexin/calreticulin
(CNX/CRT) family associate with the monoglucosylated glycans
to promote folding. They release the client when also the final
glucose is removed, but if the folding is incomplete, the glycan
becomes reglucosylated by UGGT allowing another cycle of
CNX/CRT association (Helenius and Aebi, 2004; Figure 9). Also
the mannose moieties are progressively trimmed until the gly-
can structure is eventually recognized by lectins of the EDEM
 Author's personal copy
Organelles: Structure and Function: The Endoplasmic Reticulum 163

and OS9 families and dispatched to the cytosol for proteasomal
degradation (Hosokawa et al., 2001; Christianson et al., 2008).
In this way, molecules that did not have enough time to fold
are spared from degradation, while those client proteins that
linger in the ER for too long because they fail to fold are retro-
translocated to the cytosol. Here, they are deglycosylated, ubi-
quitinated, and, finally, degraded by the proteasome, a process
referred to as ER-associated degradation (ERAD) (Werner et al.,
1996). Rarely, misfolded clients may escape ERAD and accu-
mulate instead in the ER, sometimes even forming aggregates.
Autophagy may serve as a backup removal mechanism. Yet,
persistent accumulation of misfolded ER clients often leads to
ER storage diseases (Anelli and Sitia, 2010).
In general, however, the combined efforts of the ER resi-
dent chaperones and foldases promote productive folding,
and, at the same time, ensure through their persistent binding
that immature clients are retained in the ER. Only when clients
are fully folded (and properly assembled in case of multimeric
proteins) do all chaperones release and allow exit of clients to
travel further along the secretory pathway. As such, stringent
quality control is exerted on the secretory proteome (Ellgaard
and Helenius, 2003; Anelli and Sitia, 2008).
Export from and Retrieval to the ER
Once client proteins are correctly folded, they are sorted to ER
exit sites. Some clients team up with dedicated cargo receptors
to promote this sorting process. Notable examples include the
lectin ER–Golgi intermediate compartment (ERGIC-53)
(Appenzeller et al., 1999). At ER exit sites membrane curvature
is promoted through the buildup of a coat. Key players in this

2 Movement

1 Budding 3 Docking
pH 7.1 pH 6.7

COPII coat
4 Uncoating
and fusion
ER
Golgi

COPI coat
4 Uncoating
and fusion

3 Docking
1 Budding

2 Movement

KDEL receptor Receptor-dependent cargo v-SNARE

Lumenal protein with KDEL

Receptor for lumenal cargo t-SNARE
retrieveal signal
Membrane cargo Bulk-flow cargo Tether

Figure 10 ER export and retrieval. Fully folded ER client proteins exit from the ER at ER exit sites, where they are packaged into COPII coated
vesicles. Some clients are specifically recruited for exit by cargo receptors, while other clients enter these vesicles by bulk flow. The COPII vesicles
bud from ER exit sites and travel through anterograde transport to the cis-Golgi. The vesicles dock through tethering of the two membranes and
fuse through the action of SNARES on both vesicle (v) and target (t) membranes. The cargo is released into the Golgi and the COPII coat
dissociates. Retrograde transport is facilitated by COPI coated vesicles, that let cargo receptors and KDEL receptors travel back to the ER. KDEL
receptors pick up ER chaperones and folding enzymes that display a KDEL (-like) tetrapeptide at their C-terminus.
 Author's personal copy
164 Organelles: Structure and Function: The Endoplasmic Reticulum

Enhance Reduce Apoptosis

folding folding
capacity load

ATF6 Cleaved ATF6 drives UPR targets

Spliced XBP1 drives UPR targets

Spliced XBP1 drives UPR targets that promote ERAD

IRE1
RIDD reduces mRNA levels of ER clients

IRE1 activates JNK1 and BAK/BAX pro-apoptotic pathways

eIF2α phosphorylation blocks protein synthesis

PERK
Persistent PERK mediated CHOP induction leads to apoptosis

Figure 11 Overview of how UPR pathways aim for ER homeostasis or give way to apoptosis.

process are the components of coat protein complex II
(COPII) and the GTPase Sar1 (Barlowe et al., 1994). GTP hy-
drolysis guides both the installment of the coat and the
eventual budding from the ER of COPII-coated vesicles that
contain the client proteins (Kuehn et al., 1998; Figure 10). The
vesicles then undergo anterograde transport along the secre-
tory pathway, traveling via the ERGIC, the Golgi apparatus, the
trans-Golgi network before reaching the plasma membrane as
the final destination. Through the endocytic pathway, i.e., via
endosomes to lysosomes, proteins may travel back into the
cell. The secretory and endocytic pathways therefore form a
continuous endomembrane system that is interconnected
through vesicular transport. Accordingly, almost all proteins of
the endomembrane system originate from the ER.
Specific SNARE proteins mark transport vesicles to define
their origin and they team up selectively with those SNARE
proteins that mark the designated target membrane. Rab
family members further assist in ensuring proper delivery of
vesicles. The SNAREs on the two membranes combined drive
fusion of the vesicle with the target organelle (ERGIC or Golgi)
and release of the client protein cargo into the next station
along the secretory pathway (Sudhof and Rothman, 2009;
Figure 10). While most ER clients that are subunits of multi-
meric complexes need to oligomerize first in order to exit the
ER, some assembly steps may take place in the ERGIC, as is the
case for oligomerization of immunoglobulin M (Anelli et al.,
2007). In principle, folding and assembly are thus fully com-
pleted when clients arrive in the Golgi. In downstream com-
partments of the secretory pathway, N-linked glycans often are
further modified in several ways, yielding varied complex
glycan structures. In the trans-Golgi network, some clients are
even proteolytically cleaved by furin(-like) endoproteases to
reach their mature state.
If immature client proteins untimely exit the ER, they
usually are retrieved from further stations along the secretory
pathway through retrograde transport. The ER chaperones and
foldases in small numbers do travel to the Golgi, where they
can pick up escapee clients. The chaperones carry at their
C-terminus a lysine aspartate glutamate leucine (KDEL) or
related tetrapeptide (Munro and Pelham, 1987), which is
recognized by KDEL receptors in the Golgi. The ternary client–
chaperone–KDEL receptor complex is then sorted to COPI-
coated vesicles that travel back to the ER for further client
folding attempts (Lewis and Pelham, 1992; Figure 10). The pH
gradient between the ER and Golgi contributes in regulating
transport and quality control. ERGIC-53 captures glyco-
proteins in the ER releasing them in the more acidic pH in the
Golgi, where instead the chaperone ERp44 and KDEL receptors
are activated (Wilson et al., 1993; Vavassori et al., 2013). The
ternary complex formed with clients is retrieved back to the
ER, whose neutral pH favors dissociation. It is thought that
KDEL receptors keep track of the protein flux exiting the ER
and accordingly act as signaling devices (they are structurally
and functionally similar to GPCRs), to drive adaptations in
vesicular transport capacity to and from the Golgi (Pulvirenti
et al., 2008).
ER Homeostasis
To maintain homeostasis, the ER folding capacity or efficiency
must be adjusted to the folding load, which may vary during
differentiation or due to changing environmental cues (drugs
or cofactor deficiencies). In essence, the ER perceives accu-
mulation of aberrant or orphan proteins that cannot complete
their folding schedule. The intricate sensing mechanisms that
 Author's personal copy
Organelles: Structure and Function: The Endoplasmic Reticulum
monitor load and that trigger a response to ‘ER stress’ are
collectively referred to as the UPR pathways (Ron and Walter,
2007). In metazoans, the UPR is not only key for mitigating ER
stress, but also is essential for development and maintenance
of professional secretory cells, for instance during plasma cell
differentiation (Iwakoshi et al., 2003) or in pancreatic β-cell
physiology (Harding et al., 2001).
Insufficient protein folding capacity in the ER is detected
by UPR transducers of the IRE1, PERK, and ATF6 families
(Ron and Walter, 2007). They all are transmembrane proteins
with an ER luminal stress sensing domain and a cytosolic ef-
fector domain. ATF6 proteins – which are exclusive to meta-
zoans – travel from the ER to the Golgi. Here, cleavage by a
resident protease allows the ATF6 cytosolic domain to detach,
such that it can relocate to the nucleus, where it activates
transcription of target genes (Mori, 2009). IRE1 proteins –
which are conserved in all eukaryotes – initiate the non-
conventional splicing of xbp1 mRNA (HAC1 in yeast),
whereupon the spliced mRNA is translated into another tran-
scription activator of distinct UPR target genes (Ron and
Walter, 2007). PERK – exclusive to animal cells – phos-
phorylates eIF2α, which causes translational attenuation,
thereby blocking further entry of ER clients. Paradoxically,
synthesis of some other proteins that further control the re-
sponse (e.g., ATF4 and CHOP) is enhanced when eIF2α is
phosphorylated (Ron and Walter, 2007).
UPR target genes include a full repertoire of ER chaperones
and foldases, but also enzymes for membrane lipid synthesis
that together sustain expansion of the ER and its folding
capacity. Other UPR targets are ERAD components, which
enhance the clearance of misfolded clients from the ER. IRE1
proteins may also contribute to the curtailing of the folding
load in the ER through degradation of mRNAs other than
xbp1, a process referred to as regulated Ire1-dependent decay
(Hollien and Weissman, 2006). Finally, excess or damaged ER
with its contents can be targeted for removal by autophagy
(Bernales et al., 2006). Such ‘ERphagy’ may also be key to
downsize the ER when cells are recovering from ER stress or to
sustain balanced ER expansion in professional secretory cells
(Pengo et al., 2013).
Yet, when all adaptive measures are exhausted, the UPR
induces apoptosis, for instance through persistent activation of
PERK and its downstream effector CHOP. Also IRE1 can in-
voke pro-apoptotic pathways (Ron and Walter, 2007). Mal-
adaptive ER stress responses can lie at the basis of a growing
list of pathologies. For example, excessive insulin synthesis
and a corresponding ‘overzealous’ UPR in pancreatic β-cells
lead to their demise and, hence, type 2 diabetes (Oyadomari
et al., 2002). A schematic overview of how the UPR orches-
trates ER homeostasis is shown in Figure 11.
Perspectives
Many questions remain open concerning the ER. How do cells
set and maintain an optimal ratio between ER subdomains?
What are all the distinguishing components of subdomains
and the plethora of ER contact sites? How is production in the
ER factory coordinated with the flux of proteins through the
secretory pathway? How do developmental cues determine
165
that the ER specializes in one of its many manifestations?
Which are the cues that activate and direct ERphagy? Can we
reconstruct the ER in a test tube? Can we create model cells
with a ‘tailor-made’ ER for biotechnological purposes? It is
clear that the ER has a multitude of surprising and exciting
features in store that remain to be discovered.
Acknowledgments
The authors wish to thank the Associazione Italiana della
Ricerca sul Cancro (EvA, RS), the Ministero della Salute (EvA,
RS), the Armenise–Harvard Foundation (EvA), and Telethon
(RS) for grant support.
See also: Organelles: Structure and Function: Intermediate
Compartment: A Sorting Station between the Endoplasmic Reticulum
and the Golgi Apparatus
References
Anelli, T., Ceppi, S., Bergamelli, L., et al., 2007. Sequential steps and checkpoints
in the early exocytic compartment during secretory IgM biogenesis. The EMBO
Journal 26, 4177–4188.
Anelli, T., Sitia, R., 2008. Protein quality control in the early secretory pathway.
EMBO Journal 27 (2), 315–327.
Anelli, T., Sitia, R., 2010. Physiology and pathology of proteostasis in the early
secretory compartment. Seminars in Cell & Developmental Biology 21, 520–525.
Anfinsen, C.B., 1973. Principles that govern the folding of protein chains. Science
181, 223–230.
Appenzeller, C., Andersson, H., Kappeler, F., Hauri, H.P., 1999. The lectin
ERGIC-53 is a cargo transport receptor for glycoproteins. Nature Cell Biology 1,
330–334.
Appenzeller-Herzog, C., Ellgaard, L., 2008. The human PDI family: Versatility packed
into a single fold. Biochimica et Biophysica Acta 1783, 535–548.
Barlowe, C., Orci, L., Yeung, T., et al., 1994. COPII: A membrane coat formed by
sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell 77,
895–907.
Bernales, S., McDonald, K.L., Walter, P., 2006. Autophagy counterbalances
endoplasmic reticulum expansion during the unfolded protein response. PLoS
Biology 4, e423.
Berridge, M.J., 2002. The endoplasmic reticulum: A multifunctional signaling
organelle. Cell Calcium 32, 235–249.
Blobel, G., Dobberstein, B., 1975. Transfer of proteins across membranes. I.
Presence of proteolytically processed and unprocessed nascent immunoglobulin
light chains on membrane-bound ribosomes of murine myeloma. Journal of Cell
Biology 67, 835–851.
Bulleid, N.J., Ellgaard, L., 2011. Multiple ways to make disulfides. Trends in
Biochemical Sciences 36, 485–492.
Chen, Y., Zhang, Y., Yin, Y., et al., 2005. SPD − a web-based secreted protein
database. Nucleic Acids Research 33, D169–D173.
Christianson, J.C., Shaler, T.A., Tyler, R.E., Kopito, R.R., 2008. OS-9 and GRP94
deliver mutant alpha1-antitrypsin to the Hrd1-SEL1L ubiquitin ligase complex for
ERAD. Nature Cell Biology 10, 272–282.
Cox, J.S., Shamu, C.E., Walter, P., 1993. Transcriptional induction of genes
encoding endoplasmic reticulum resident proteins requires a transmembrane
protein kinase. Cell 73, 1197–1206.
Eletto, D., Dersh, D., Argon, Y., 2010. GRP94 in ER quality control and stress
responses. Seminars in Cell & Developmental Biology 21, 479–485.
Ellgaard, L., Helenius, A., 2003. Quality control in the endoplasmic reticulum. Nature
Reviews Molecular Cell Biology 4, 181–191.
Endo, M., 1977. Calcium release from the sarcoplasmic reticulum. Physiological
Reviews 57, 71–108.
English, A.R., Voeltz, G.K., 2013. Rab10 GTPase regulates ER dynamics and
morphology. Nature Cell Biology 15, 169–178.
 Author's personal copy
166 Organelles: Structure and Function: The Endoplasmic Reticulum
Fagone, P., Jackowski, S., 2009. Membrane phospholipid synthesis and
endoplasmic reticulum function. Journal of Lipid Research 50 (Suppl.),
S311–S316.
Galat, A., 2003. Peptidylprolyl cis/trans isomerases (immunophilins): Biological
diversity − targets − functions. Current Topics in Medicinal Chemistry 3,
1315–1347.
Gilmore, R., Blobel, G., Walter, P., 1982. Protein translocation across the
endoplasmic reticulum. I. Detection in the microsomal membrane of a
receptor for the signal recognition particle. Journal of Cell Biology 95, 463–469.
Goder, V., Spiess, M., 2001. Topogenesis of membrane proteins: Determinants and
dynamics. FEBS Letters 504, 87–93.
Goldenberger, R.F., Epstein, C.J., Anfinsen, C.B., 1963. Acceleration of reactivation
of reduced bovine panreatic ribonuclease by a microsomal system from rat liver.
Journal of Biological Chemistry 238, 628–635.
Goyal, U., Blackstone, C., 2013. Untangling the web: Mechanisms underlying
ER network formation. Biochimica et Biophysica Acta 1833, 2492–2498.
Harding, H.P., Zeng, H., Zhang, Y., et al., 2001. Diabetes mellitus and exocrine
pancreatic dysfunction in perk-/- mice reveals a role for translational control in
secretory cell survival. Molecular Cell 7, 1153–1163.
Helenius, A., Aebi, M., 2004. Roles of N-linked glycans in the endoplasmic
reticulum. Annual Review of Biochemistry 73, 1019–1049.
Helle, S.C., Kanfer, G., Kolar, K., et al., 2013. Organization and function
of membrane contact sites. Biochimica et Biophysica Acta 1833,
2526–2541.
Hollien, J., Weissman, J.S., 2006. Decay of endoplasmic reticulum-localized mRNAs
during the unfolded protein response. Science 313, 104–107.
Hosokawa, N., Wada, I., Hasegawa, K., et al., 2001. A novel ER alpha-
mannosidase-like protein accelerates ER-associated degradation. EMBO Reports
2, 415–422.
Hu, J., Shibata, Y., Zhu, P.P., et al., 2009. A class of dynamin-like
GTPases involved in the generation of the tubular ER network. Cell 138,
549–561.
Hurtley, S.M., Bole, D.G., Hoover-Litty, H., Helenius, A., Copeland, C.S., 1989.
Interactions of misfolded influenza virus hemagglutinin with binding protein
(BiP). Journal of Cell Biology 108, 2117–2126.
Iwakoshi, N.N., Lee, A.H., Vallabhajosyula, P., et al., 2003. Plasma cell
differentiation and the unfolded protein response intersect at the transcription
factor xbp-1. Nature Immunology 4, 321–329.
Keenan, R.J., Freymann, D.M., Stroud, R.M., Walter, P., 2001. The signal
recognition particle. Annual Review of Biochemistry 70, 755–775.
Klopfenstein, D.R., Klumperman, J., Lustig, A., et al., 2001. Subdomain-
specific localization of climp-63 (p63) in the endoplasmic reticulum is
mediated by its luminal alpha-helical segment. Journal of Cell Biology 153,
1287–1300.
Kopito, R.R., Sitia, R., 2000. Aggresomes and Russell bodies. Symptoms of cellular
indigestion? EMBO Reports 1, 225–231.
Kuehn, M.J., Herrmann, J.M., Schekman, R., 1998. CopII-cargo interactions
direct protein sorting into ER-derived transport vesicles. Nature 391, 187–190.
Lang, S., Benedix, J., Fedeles, S.V., et al., 2012. Different effects of sec61
alpha, sec62 and sec63 depletion on transport of polypeptides into the
endoplasmic reticulum of mammalian cells. Journal of Cell Science 125,
1958–1969.
Lewis, M.J., Pelham, H.R., 1992. Ligand-induced redistribution of a human KDEL
receptor from the Golgi complex to the endoplasmic reticulum. Cell 68, 353–364.
Lin, S., Sun, S., Hu, J., 2012. Molecular basis for sculpting the endoplasmic reticulum
membrane. International Journal of Biochemistry & Cell Biology 44, 1436–1443.
Liu, B., Yang, Y., Qiu, Z., et al., 2010. Folding of toll-like receptors by the HSP90
paralogue gp96 requires a substrate-specific cochaperone. Nature
Communications 1, 1–10.
Mayer, M.P., Bukau, B., 2005. HSP70 chaperones: Cellular functions and molecular
mechanism. Cellular and Molecular Life Sciences 62, 670–684.
Meldolesi, J., Pozzan, T., 1998. The endoplasmic reticulum Ca2 þ store: A view
from the lumen. Trends in Biochemical Sciences 23, 10–14.
Mori, K., 2009. Signalling pathways in the unfolded protein response: Development
from yeast to mammals. Journal of Biochemistry 146, 743–750.
Mori, K., Ma, W., Gething, M.J., Sambrook, J., 1993. A transmembrane protein with
a cdc2 þ /CDC28-related kinase activity is required for signaling from the ER to
the nucleus. Cell 74, 743–756.
Munro, S., Pelham, H.R., 1987. A C-terminal signal prevents secretion of luminal
ER proteins. Cell 48, 899–907.
Naon, D., Scorrano, L., 2014. At the right distance: ER-mitochondria
juxtaposition in cell life and death. Biochimica et Biophysica Acta 1843,
2184–2194.
Novick, P., Field, C., Schekman, R., 1980. Identification of 23 complementation
groups required for post-translational events in the yeast secretory pathway. Cell
21, 205–215.
Orsi, A., Sitia, R., 2007. Interplays between covalent modifications in the
endoplasmic reticulum increase conformational diversity in nascent prion protein.
Prion 1, 236–242.
Oyadomari, S., Araki, E., Mori, M., 2002. Endoplasmic reticulum stress-mediated
apoptosis in pancreatic beta-cells. Apoptosis 7, 335–345.
Palade, G.E., Porter, K.R., 1954. Studies on the endoplasmic reticulum. I.
Its identification in cells in situ. Journal of Experimental Medicine 100,
641–656.
Palade, G.E., Siekevitz, P., 1956. Liver microsomes; an integrated morphological
and biochemical study. Journal of Biophysical and Biochemical Cytology 2,
171–200.
Pengo, N., Scolari, M., Oliva, L., et al., 2013. Plasma cells require autophagy
for sustainable immunoglobulin production. Nature Immunology 14,
298–305.
Praissman, J.L., Wells, L., 2014. Mammalian O-mannosylation pathway:
Glycan structures, enzymes, and protein substrates. Biochemistry 53,
3066–3078.
Pulvirenti, T., Giannotta, M., Capestrano, M., et al., 2008. A traffic-activated Golgi-
based signalling circuit coordinates the secretory pathway. Nature Cell Biology
10, 912–922.
Ron, D., Walter, P., 2007. Signal integration in the endoplasmic reticulum unfolded
protein response. Nature Reviews. Molecular Cell Biology 8, 519–529.
Schuldiner, M., Metz, J., Schmid, V., et al., 2008. The GET complex
mediates insertion of tail-anchored proteins into the ER membrane. Cell 134,
634–645.
Shen, Y., Meunier, L., Hendershot, L.M., 2002. Identification and characterization of
a novel endoplasmic reticulum (ER) DnaJ homologue, which stimulates ATPase
activity of BiP in vitro and is induced by ER stress. Journal of Biological
Chemistry 277, 15947–15956.
Siekevitz, P., Palade, G.E., 1960. A cytochemical study on the pancreas of the
guinea pig. 5. In vivo incorporation of leucine-1-C14 into the chymotrypsinogen
of various cell fractions. Journal of Biophysical and Biochemical Cytology 7,
619–630.
Steel, G.J., Fullerton, D.M., Tyson, J.R., Stirling, C.J., 2004. Coordinated activation
of HSP70 chaperones. Science 303, 98–101.
Stefan, C.J., Manford, A.G., Emr, S.D., 2013. ER-PM connections: Sites of
information transfer and inter-organelle communication. Current Opinion in Cell
Biology 25, 434–442.
Sudhof, T.C., Rothman, J.E., 2009. Membrane fusion: Grappling with SNARE and
SM proteins. Science 323, 474–477.
Tu, B.P., Weissman, J.S., 2004. Oxidative protein folding in eukaryotes: Mechanisms
and consequences. Journal of Cell Biology 164, 341–346.
Tyson, J.R., Stirling, C.J., 2000. Lhs1 and Sil1 provide a lumenal function that is
essential for protein translocation into the endoplasmic reticulum. EMBO Journal
19, 6440–6452.
Udenfriend, S., Kodukula, K., 1995. How glycosylphosphatidylinositol-
anchored membrane proteins are made. Annual Review of Biochemistry 64,
563–591.
Vance, D.E., Choy, P.C., Farren, S.B., Lim, P.H., Schneider, W.J., 1977. Asymmetry
of phospholipid biosynthesis. Nature 270, 268–269.
Vavassori, S., Cortini, M., Masui, S., et al., 2013. A pH-regulated quality control
cycle for surveillance of secretory protein assembly. Molecular Cell 50,
783–792.
Veratti, E., 1961. Investigations on the fine structure of striated muscle fiber read
before the Reale Istituto Lombardo, 13 march 1902. Journal of Biophysical and
Biochemical Cytology 10 (Suppl. 4), 1–59.
Voeltz, G.K., Prinz, W.A., 2007. Sheets, ribbons and tubules - how organelles get
their shape. Nature reviews. Molecular Cell Biology 8, 258–264.
Voeltz, G.K., Prinz, W.A., Shibata, Y., Rist, J.M., Rapoport, T.A., 2006. A class of
membrane proteins shaping the tubular endoplasmic reticulum. Cell 124,
573–586.
Walter, P., Ibrahimi, I., Blobel, G., 1981. Translocation of proteins across the
endoplasmic reticulum. I. Signal recognition protein (SRP) binds to in-vitro-
assembled polysomes synthesizing secretory protein. Journal of Cell Biology 91,
545–550.
Wang, F., Chan, C., Weir, N.R., Denic, V., 2014. The GET1/2 transmembrane
complex is an endoplasmic-reticulum membrane protein insertase. Nature 512,
441–444.
Werner, E.D., Brodsky, J.L., McCracken, A.A., 1996. Proteasome-dependent
endoplasmic reticulum-associated protein degradation: An unconventional route
 Author's personal copy
Organelles: Structure and Function: The Endoplasmic Reticulum
to a familiar fate. Proceedings of the National Academy of Sciences of the United
States of America 93, 13797–13801.
Wilson, D.W., Lewis, M.J., Pelham, H.R., 1993. pH-dependent binding of KDEL to
its receptor in vitro. Journal of Biological Chemistry 268, 7465–7468.
Xu, N., Zhang, S.O., Cole, R.A., et al., 2012. The FATP1-DGAT2 complex facilitates
lipid droplet expansion at the ER-lipid droplet interface. Journal of Cell Biology
198, 895–911.
van der Zand, A., Gent, J., Braakman, I., Tabak, H.F., 2012. Biochemically distinct
vesicles from the endoplasmic reticulum fuse to form peroxisomes. Cell 149,
397–409.
Zimmermann, R., Mollay, C., 1986. Import of honeybee prepromelittin into the
endoplasmic reticulum. Requirements for membrane insertion, processing, and
sequestration. Journal of Biological Chemistry 261, 12889–12895.
Further Reading
Aebi, M., Bernasconi, R., Clerc, S., Molinari, M., 2010. N-glycan structures:
Recognition and processing in the ER. Trends in Biochemical Sciences 35,
74–82.
View publication stats
167
Anfinsen, C.B., 1972. Studies on the Principles that Govern the Folding of Protein
Chains. Nobel Lecture, December 11, 1972.
Blobel, G., 1999. Protein Targeting. Nobel Lecture, December 8, 1999.
Braakman, I., Hebert, D.N., 2013. Protein folding in the endoplasmic reticulum. Cold
Spring Harbor Perspectives in Biology 5, a013201.
Brodsky, J.L., 2012. Cleaning up: ER-associated degradation to the rescue. Cell 151,
1163–1167.
Ellgaard, L., Molinari, M., Helenius, A., 1999. Setting the standards: Quality control
in the secretory pathway. Science 286, 1882–1888.
English, A.R., Voeltz, G.K., 2013. Endoplasmic reticulum structure and
interconnections with other organelles. Cold Spring Harbor Perspectives in
Biology 5, a013227.
Palade, G.E., 1974. Of Protein Secretion. Nobel Lecture, December 12, 1974.
Schekman, R.W., 2013. Genetic and Biochemical Dissection of the Secretory
Pathway. Nobel Lecture, December 7, 2013.
Walter, P., Ron, D., 2011. The unfolded protein response: From stress pathway to
homeostatic regulation. Science 334, 1081–1086.