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Coating and uncoating of vessicles

Accessory proteins fuse the bilayers

ORGANIZATION OF MEMBRANE DOMAINS

Arf GTPase: COPI @ Golgi -> ER


Clatherin @ Golgi -> ER
SarI GTPase: COPII @ ER
Reversible: GTP -> GDP
release mbr
-> disassembly

Amphipathic helix associates with lipid bilayer

Vessicle formation at
mbr regions already curved
Rims golgi cisternae
cholesterol:lipid domains

Uncoating of vessicles as soon as released


Expose targeting information

Select target membrane


surface markers
complementary R
Rab and Snare
Rab: m-GTPase
Snare mediate mbr fusion

Snare bring mbrs close and then spontaineously fuse


Lipid bilayer -> 1.5 -> lipid exchange and fusioin

Similar to snare fusion mechanism

Infleunze virus diff mech


R mediate endocytosis
-> endosome, low pH
fusion prot conf delta

Retromer
Coincidence R

Endosome to golgi

Some cargo receptors are


lectins, recognize sugars

Recall prots must be folded


immune system
infection -> proteosome
MDR transporters ER
peptides-MHC -> cell surf
T-lymphocytes

ER <-> Golgi
Bidirectional
ER -> in COPII vesicles
@ ER exit sites
no ribosomes
Golgi
carbohydrate synthesis
N- and O-linked
sort and dispatch from ER ->
oligosaccharides are target seq
for specific transport vesicles

Homotypic vessical fusion after coat loss: rab/snare

ER sorting signals
KKXX (mbr) & KDEL (soluble ER resident)
pH-dependent affinity

ER

PM

Tubular connections
between stacks
cisterna contiguous

Diff function diff prot/enzymes


Process N-linked oligosacchs
CGN sorting: oligo phosphorylation -> target (lysosome)
Cis cisterna: remove Man
Medial cisterna: remove Man add GlcNAc
Trans cisterna: add Gal:NANA
TGN: sulfation tyrosines and carbohydrates -> sorting

Reduced osmium

dinucleotide ppase
Acid ppase

Goblet cells
Intestinal epithelium
Mucus secretion
mucins (o-link)
Polysaccharide rich
lots golgi

ER

Targeted glycosylation

Golgi

Summary

Rab/Snare interactions at target


Mbr domain organization
Retrograde transport
Microtubule Road maps
Viral entry

Retromer
Coincidence R

Endosome to golgi

Lysosomes

pH 4-5

pH 6

pH 7.2

pH 6.5-6.7

Lysosomes: pathway convergence


materials from ER enzymes
zymogens
materials from outside
endocytosis
materials from autophagocytosis
phagocytosis
Cargo signal
mannose 6-phosphate, N-link
Cargo receptor
mannose 6-phosphate

Mannose-6-P

I-cell disease

Endocytosis
Pinocytosis
Phagocytosis
Autophagocytosis

Endocytosis

pH 4-5

pH 6

pH 7.2

pH 6.5-6.7

http://www.youtube.com/watch?v=1cuaS2QKobA&feature=related

Endocytosis
Pinocytic vesicle -> clatherin coated pits

Caveolae vs clatherin

Cholesterol

Cholesterol

Steroid hormones

Vitamin D3 synthesis, ricketts


Cholesterol ester

Fuse with lysosome


Release cholesterol

<-

Fuse with endosome low pH


Release receptor

Patients with Cholesteryl Ester Storage Disease


typically have elevated cholesterol and
triglyceride levels (type II hyperlipidemia) and
may present at an early age with the following
liver abnormalities:
Unexplained hepatomegaly
infection
direct toxicity
hepatic tumours
metabolic disorder
Elevated transaminases (ALT,
AST,AP,bilirubin)
Unexplained fatty liver
Progressive and/or unexplained
chronic liver disease

Vessicle fates
transcytosis
recycling
degradation
storage

Fates of R that have been endocytosed


2. Caveolae caveolins
SV40 virus

3. EGF receptor
downregulation
1. LDL R
1. Transferrin R
iron

ubiquitin

Transferrin R

-> ER, misfold

Ricin

Sorting ESCRT complx


4 prot complx
GPI-PI(3)(3.5)P mediated
Ubiquitin tag

Transcytosis
Fc receptor

Storage of R
In recycling endosomes

Insulin hexamer

Mobilize when needed


Glucose transporter
Insulin stimulated fusion

Two distinct endosome


compartments
If cargo has no signal
For recycle or transcytosis
-> lysosome

Exocytosis

Hormeones, neurotransmitors

Mannose-6-PO4

ATP-driven H+ pumps acidify all endo/exo vesicles

200-400 fold conc

Pancreatic B cell
insulin

Secretory vesicle

excite

acetylcholine

glutamate

serotonin

GABA
glycine
inhibit

Hemoglobin S

Hemoglobin Tetramer

G-Protein Coupled
Receptor! !
N!

Effector!
Ligand!

7!
1!

Lipid Anchor!

C!

2!

6!

Cytoplasm!

5!
3!

GTP!

GDP!

4!

Cell!
Membrane!

7-transmembrane!
-helix protein!