 IntracellularCompartments and

Protein Sorting
The major intracellular compartments of an animal cell
An electron
micrograph of part of a
live cell seen in cross
section
Topological relationships between compartments of the
secretory and endocytic pathways in a eucaryotic cell
A schematic roadmap of
protein traffic
Red: gated transport
Blue: transmembrane
transport
Green: vesicular transport
Vesicle budding and fusion
during vesicular transport
Two ways in which a sorting signal can be built into a
protein
a. Signal sequence
b. Signal patch
 The transport of molecules between
the nucleus and the cytosol
The nuclear envelope

Nuclear envelope of the typical

mammalian cell contains 3000-4000
pore complexes.
The arrangement of nuclear pore complexes in the nuclear
envelope
Column-stubaste podjedinice (narandžasta)
Annular-okrugle podjedinice (zelena)
Lumenal-lumenalne podjedinice (plava)
Ring-prstenaste podjedinice (žuta)
Possible paths for free diffusion through the nuclear pore
complex

9 nm

Small molecules < 5000 Da diffuse in

so fast that the nuclear envelope can be
considered to be freely permeable to them.
Protein >60 000 Da is not able to enter nucleus.
The function of a nuclear
localization signal
a. Nuclear localization
signal: NLS (rich in the
positively charged amino
acids Lys and Arg)
b. Nuclear export signal:
NES
Nuclear import receptors: soluble cytosolic proteins that
bind both to the nuclear localisation signal on the protein and
to nucleoporins.

Nucleoporins: some of them form fibrils that extend into

Cytosol from the rim of the nuclear pore complex.
The Ran GTPase Drives Directional Transport Through
Nuclear Pore Complexes

 The import of nuclear proteins through the

pore complex concentrates specific proteins
in the nucleus, thereby increasing order in
the cell, which must consume energy.
 The energy is thought to be provided by the

hydrolysis of GTP by the monomeric GTPase

Ran. Ran is found in both the cytosol and the
nucleus, and it is required for both the
nuclear import and export systems.
 The endoplasmic reticulum
The rough ER
Free and membrane-bound ribosomes
The Isolation of purified rough and smooth microsomes from
the ER
The signal hypothesis
The signal-recognition particle (SRP)
How ER signal sequences and SRP direct ribosomes to the ER
membrane
The Polypeptide Chain Passes Through an Aqueous Pore in the
Translocator

 The translocator, called the Sec61 complex,

consists of three or four protein complexes,
each composed of three transmembrane
proteins, that assemble into a donutlike
structure.
 The pore is a dynamic structure that opens

only transiently when a ribosome with a

growing polypeptide chain attaches to the ER
membrane.
Three ways in which protein translocation can be driven
through structurally similar translocators
A model for how a soluble protein is translocated across the
ER membrane
How a single-pass transmembrane protein with a cleaved ER
signal sequence is integrated into the ER membrane
Integration of a single-
pass membrane protein
with an internal signal
sequence into the ER
membrane
Integration of a double-pass membrane protein with an internal
signal sequence into the ER membrane
The insertion of the multipass membrane protein rhodopsin into
the ER membrane
The asparagine-linked (N-linked)
precursor oligosaccharide that is
added to most proteins in the
rough ER membrane
Protein glycosylation
in the rough ER

oligosaccharyl
transferase
enzyme adds
oligosaccharide
to asparagine
The role of N-linked glycosylation in ER protein folding
Calnexin: membrane-bound chaperone protein
Calreticulin: soluble chaperone protein
The export and degradation of misfolded ER proteins
The attachment of a GPI (glycosylphosphatidyl-
inositol) anchor to a protein in the ER
The role of phospholipid translocation in lipid bilayer
synthesis
 Summary
Nucleus translocation, NLS, NES,
nuclear pore complex, Ran-GTP
Endoplasmic reticulum, rough ER,

smooth ER,
SRP, soluble and membrane proteins in

ER,
Glycosylation in ER, folding,
Membrane lipid bilayer assembly