CBG Lecture 36: Vesicular Traffic Flashcards
what are the main experiments that led to the concept of trafficking
pulse chain:radioactivity
GFP - encoded by cDNA then attach GFP tag to viral glycoprotein VSVG - use glycoprotein as this will undergo postranslational modification - in Golgi
why do proteins move
prevent depletion of organelle-resident proteins
prevent depletion of lipids
what is KDEL sequence
Golgi to ER signalling sequence
where must proteins destined for the signalling pathway go
to the RER
via what do proteins enter the RER
via SRP signal peptide recognition partice
where does synthesis and modification of proteins occur
RER
where does synthesis of lipids occur
SER
where does calcium storage occur
sarcoplasmic reticulum in muscle cells
how does vesicular transport betwen compartments occur
budding and fusion
what are the different types of protein vesicle coats
clathrin
COPI
COPII
what do protein coats do
organise vesicular traffic and give shape to vesicles and concentrate membrane proteins in a specialised patch
what is a vesicle coat
a small membrane bounded organelle with a cage of proteins (coat) on its cytosolic surface.
formed by piclking off of a coated region of membrane - coated pit
where do clathrin coats mediate transport to
golgi
PM
where do COPI coats mediate transport to
ER and Golgi cisternae
what are the major components of clathrin coated vesicles
clathrins and adaptins
each clathrin unit has 3 large heavy and 3 light pp chains that form a triskelion
where is triskelion found
what is it made up of
triskelion found in clathrin coated vesicles
made up of 3 large heavy and 3 light pp chains
alpha zigzag and beta propellor- 36 triskelions make up a hexagonal clathrin cage
what can isolated triskelions do
self assemble sponatenously
what are adaptin proteins
requierd to bind the clathrin coat to membrane and to trap transmembrane receptors that capture soluble cargo molecules
outline the structure of clathrin
alpha zigzag
beta propeller makes the triskelion
36 triskelions make up a hexagonal clathrin cage
how many triskelions make up a hexagonal clathrin cage
36
discuss assembly and disassembly of clathrin coat
assembly of coat thought to induce curvature into membrane, which leads to formation uniformly sized coated buds
- adaptins bind both clathrin triskelions adn membrane bound cargo receptors, thereby mediating the selective recruitment of both membrane and cargo molecules into the vesicle
- ARF GTPase initiastes assembly
- dynamin pinches off vesicle
- ARF switching and Hsc70 ATP hydrolysis triggers uncoating of vlathrin
which enzymes initiate assembly of clathrin coated vesicles
ARF GTPase
which enzymes trigger uncoating of clathrin coated vesicles
ARF switching
and Hsc70 ATP hydrolysis
what are the four adaptor proteins present in clathrin coated vesicles
AP1 AP3 AP2 GGA
which adaptor proteins are involved in movement from TGN trans golgi network to endosome
AP1 or GGA
which adaptor proteins are involved in movement from TGN to PM
AP2