BCH3021 Lecture Notes - Lecture 20: Retromer, Ldl Receptor, Phosphatidylinositol

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25 May 2018
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Lecture 20 Secretory Vesicle Structure and Trafficking
Proteins are transported between organelles in transport vesicles
There are different types of vesicles with different destinations
Vesicle formation involves polymerization of coat proteins
Vesicles move along microtubules
Docking and fusion of vesicles is receptor-mediated
Four Classes of Transporters in the Secretory Pathway
COP II ER to Golgi
COP I Golgi to ER
Clathrin
o TGN to endosomes
o Surface to endosomes
o TGN to basolateral membrane
Retromer endosome to TGN
Unknown TGN to surface in non-polarized cells
Vesicles are Defined by Coat Structure
Vesicles are formed by reversible polymerization of coat proteins and other
components on a membrane surface
The type of vesicle can thus be defined by its coat protein
Clathrin-coated vesicles move between TGN, surface, early endosome
Coatomer-coated vesicles move between ER and Golgi
o COP II vesicles move from ER to Golgi
o COP I vesicles move from Golgi to ER
Retromer coated vesicles move between endosomes and TGN
Targeting of Cargo Proteins to Vesicles Requires Information
Specific sorting signals on cargo proteins direct them into correct vesicles
Signals are short amino acid motifs in cytoplasmic domain of cargo protein
interact with adaptors or vesicle coat proteins
A cargo protein may also be a receptor that captures a soluble molecule
lacking a signal e.g. LDL receptor captures LDL
Steps in Vesicle Trafficking
Problems cell faces: having proteins in
membrane bound compartment
o Crossing two membranes
Solves: pinching off membrane
spherical transport vehicle
Distinguishing organelles: Phosphoinositides
Inositol phospholipids make up <10% of
membranes
Membranes vary in physiological
properties
Rapidly converted to phosphoinositides
by kinases (enzymes that add phosphates
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to head group), and modified by phosphatases (enzymes that remove
phosphates from head group)
Proteins can bind to head groups of specific phosphoinositides
Phosphoinositides assist in coat assembly and act as markers of organelle
identity
Molecular Mechanisms of Vesicular Traffic
How are transport vesicles formed?
How do vesicles move?
What dictates vesicles specificity?
How does vesicle fuse with target organelle?s
Phosphoinositides act as organelle identifiers in vesicular
trafficking
Answers how vesicles know where to go
Different organelles have different sets of
phosphoinositides
Pls can be inter-converted e.g. PI(4)P can be
converted into PI(4,5)P2 by a kinase
During vesicle formation adaptor or coat proteins
bind to specific phosphoinositides controlling where
and what type of coats assemble
Rab Proteins
Can use rab to fingerpint different
structures/organelles within secretory pathway
Lysosomes: Rab7
>60 Rab proteins in mammals
Every secretory or endocytic organelle or vesicle has at least one Rab type on
its surface
Different vesicles have different Rabs explain targeting specificity
Features of Budding and Vesicle Formation
GTP binding protein regulates the rate of vesicle
formation (molecular switch)
Adaptor proteins select the cargo protein by
recognizing a signal motif
Coat and adaptor proteins polymerize on cytoplasmic
face of budding vesicle and deform the membrane
Sanction specific vesicles
Example: initiation of vesicle formation (COP II)
Binding of inactive Sart1-GDP (soluble) to Sar1-
guanine-nucleotide exchange factor (initiator in membrane) causes release
of GDP and binding of GTP causes conformational change in Sar1
Helical tail of Sar1 exposed and mediates membrane binding
Membrane bound Sar1 binds to coat proteins (sec23/24) promotes
assembly of coat proteins into a cage around budding membrane
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