13.

VESICULAR TRAFFIC
TRANSPORT FROM THE ER TO GOLGI APARATUS
• Proteins that are translocated to the
ER are then transported to the Golgi
Apparatus by a protein-coated vesicle
• Specific proteins are required to form
a Membrane-Enclosed Vesicle from
the small amount of ER membrane
and its contents (pinching)


Types of Coated Vesicles
• Clathrin coats vesicles as they exit
from Golgi Apparatus
• COPI retrograde vesicles (Golgi to ER)
o Vesicles that bud off from the vesicle tubular cluster
• COPII anterograde vesicles (ER to Golgi)


1. COPII-COATED VESICLES
• Sar1-GDP binds to the Sar1-GEF in the ER membrane
o This induces a release of GDP and exchange for GTP
• GTP causes of a conformational change where the hydrophobic tail is
exposed and binds to the cytosolic leaflet of the ER membrane
o This initiates a membrane curvature
• Sar1-GTP recruits 2 COPII adaptor proteins (Sec23/24)

o These adaptor proteins form the inner coat

• Coat proteins recruit cargo receptor
• A complex of two additional COPII proteins (Sec13/31) forms the outer
shell of the coat
o This complex forms a cage structure
2. BUDDING OF THE COPII VESICLE
• Buds out from a specialized region called the ER Exit Site (Regions that
lack in ribosomes)
• Soluble cargo proteins in the ER lumen have exit signals that attaches
them to the transmembrane cargo receptor
o This process can be selective or can happen by default
• Proteins without exit signals
can also enter the vesicle
(ER-resident proteins)
• Proteins that properly
folded are concentrated in
the ER inside the vesicle
3. SNARE PROTEINS AND TARGET SPECEFICITY
• Soluble NFS Attachment Protein Receptor
• Surface markers on vesicles that identifies cargo types & bind to
complementary receptors
o i.e. t-SNARE to v-SNARE
• Allows for fusion of incoming vesicles to the Golgi Apparatus
• v-SNARE: Vesicular Membrane SNARE
• t-SNARE: Target Membrane SNARE
o When v and t associates with each other, they form a trans-SNARE
complex to mediate membrane fusion of the two apposed bilayer
• Rab-GTP will allow the vesicle to associate with a tethering protein called
Rab effector that helps dock and bring closer to the target membrane to
form the trans-SNARE complex
• trans-SNARE complex provides energy for the fusion of the vesicle to the
membrane of the Golgi Apparatus

4. DOCKING AND SNARE DISSOCIATION

• SNAREs mediate docking and membrane fusion
• NFS (ATPase) “pries” SNARE apart so they can be used again for later
o NSF hydrolyzes ATP (with accessory proteins)
5. HOMOTYPIC MEMBRANE
• Budded vesicles from the same compartment fuses with each other
o Heterotypic is the fusion of membranes from different compartment
• Requires a set of matching SNAREs
o Both contributing t-SNAREs and v-SNAREs (symmetrical interaction)
• Addition of adaptor proteins, NSF and ATP pries apart of the trans-SNARE
complex
• Recognition of SNAREs from different vesicles leads homotypic fusion

PATHWAY
• NSF pries apart identical v-SNAREs & t-SNAREs in both membranes
• Separated matching SNAREs on adjacent identical membrane interact,
leading to membrane fusion and the formation of a continuous
compartment
o This leads to the formation of a Vesicular Tubular Cluster
• Compartment grows by further homotypic fusion
6. VESICULAR TUBULAR CLUSTER
• Formed by the homotypic fusion of COPII Vesicles
• Function as a transport compartment to the Cis Golgi
• Travel along the MT and compartment becomes the cargo
o Transports large structures from one compartment to another
o Transported by Kinesin along a MT

7. TRANSPORT THROUGH THE GOLGI

• As proteins move through the Golgi Apparatus, they are modified through
series of biochemical reactions
• These biochemical modifications help sort the proteins to direct them to
their appropriate destinations

8. A GOLGI “STACK”
• Series of membrane-enclosed compartments called cisternae linked by
tubular connections forming a single complex
• Materials enter from the Cis Face
• Cis Golgi Network (CGN) and trans Golgi network (TGN), find them
based on their biochemistry, different complement of enzymes, proteins
would be modified differently
9. GLYCOSYLATION
• Addition of one or more sugars on protein (or lipid)
• N-linked glycosylation: NH2 on side chain of Asparagine
o Produces to two types of oligosaccharide: Complex and High-mannose
• O-linked glycosylation: OH on side chain of Serine/Threonine

10. FUNCTIONAL COMPARTMENTALIZATION

• Trans Cisterna: Glycosylation usually complete
• Trans Golgi Network (TGN): Segregation of glycosylated proteins
• Removal of Mannose (glycosylated in the ER) occurs cis and medial
cisternae
o Important for sorting protein in the Plasma Membrane

11. 2 MODELS FOR PROTEIN TRANSPORT BETWEEN

CISTERNAE
(A) Dynamic (Cisternal Maturation Model)
• Views Golgi Cisternae as a dynamic structure
• New cisternae continually form as vesicular tubular clusters arrives and
progressively matures as medial cisternae and then trans cisternae
• Cisternae moves through the Golgi Stack with its cargo proteins in the
lumen

(B) Static (Vesicle Transport Model)

• Cisternae does not mature and remain fixed
• Forward-Moving Transport Vesicles bud from one cisternae to the next in a
cis-trans direction
• Vesicles are the ones moving through the cisternae and unload part of its
cargo in the different parts of the Golgi Apparatus


12. TRANSPORT FROM THE GOLGI ONWARD

• Proteins emerge from the Golgi and are then sent to their appropriate
destinations
• Transport vesicles carry lipid soluble proteins and lipids bound for the
Plasma Membrane and water soluble proteins for exocytosis

13. 2 SECRETORY PATHWAY

I. Constitutive Pathway
• Constitutive Secretory Pathway operates in all cell (conventional
pathway)
• Many soluble proteins that secreted from the cell by this pathway
• Pathway supplies the PM with membrane lipids and proteins

II. Regulated Pathway

• Specialized secretory cells have a Regulated Secretory Pathway
• Proteins from the TGN packaged into secretory vesicles and remain
concentrated and
stored in the
vesicles until an
unknown
signaling pathway
stimulates the
secretion
14. SECRETORY VESICLES
I. Formation
• Segregation and concentration of secretory proteins occurs by:
o Progressive acidification of vesicle lumen (by V-Type ATPase)
o Removal of membrane and lumen material (recycle) from vesicle by
Clathrin formation
• Clathrin coat is initially recruited on a bulky vesicle which removes material
away to form a smaller vesicle

II. Release
• Release of vesicular material occurs with docking
• Vesicles travel to plasma membrane
o By axonal transport for neurons
• Waits to receive an extracellular signal (chemical or electrical)
2+
o Increase in cytosolic [Ca ] could induce a release
• Content is then discharged to the cell exterior
• Fusion of the vesicle to the PM temporarily increases PM surface area
• Proteins that are components of the vesicle are either degraded or recycled