chapter 13 intracellular vesicular traffic 張學偉 助理教授
TRANSCRIPT
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Chapter 13
• Intracellular Vesicular Traffic
張學偉 助理教授
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The molecular mechanisms of membrane transport and the
maintenance of compartmental diversity
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GFP-FUSION PROTEINS HAVE REVOLUTIONIZED THE STUDY OF INTRACELLULAR TRANSPORT
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There are various types of coated vesicles
Each type is used for different transport steps in the cell.
Mediate transportfrom Golgi & fromplasma membrane
Mediate transportfrom ER & Golgi
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The assembly of a clathrin coat drives vesicle formation
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Major coat protein: clathrin & adaptin
There are at least four types of adaptins, each specific for a different set of cargo receptor.
by charperone (hsp70)
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Both the pinching-off and uncoating of coated vesicles are
regulated processes
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Not all transport vesicles are spherical
Various size & shape
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Monomeric GTPase control coat assembly
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SNARE proteins and targeting GTPases guide membrane transport
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Trans-SNARE complex
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Interacting SNAREs need to be pried apart before they can function again
cytosol
Cycle
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Rab proteins (monomeric GTPase) help ensure the specificity of vesicle docking
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Rab facilitate docking of transport vescicles.
cycleRab &its effectorStructure vary greatly
Function common(1.concentrate & tethervesicle near target site2. Trigger release of SNARP control protein)
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SNARE may mediate membrane fusion
Process for SNARE concentrate in membrane fusion
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Viral fusion proteins and SNAREs may use similar strategies
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Transport from the ER through the golgi apparatus
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Cargo Recruitment
ER exit site
50nm vesicle > 200 mem protein
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Only proteins that are properly folded and assembled can leave the ER
ER resident protein
Incomplete
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CF (Cystic fibrosis)
Defect in Cl- transport
This is not because the mutation inactivated the protein, But because the active protein is discarded before it reaches the plasma membrane.
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Transport from the ER to the Golgi apparatus is mediated by vesicular tubular clusters.
Heterotypic fusion
Homotypic memb fusion is not restricted to form VTC.
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The structure formed when ER-derived vesicles fuse with one another are called.
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ER retrival signal
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The retrieval pathway to the ER uses sorting signals
Lys-Asp-Glu-Leu (KDEL)
KKXX at c-terminal end direct interact with COPI coat
Short retrieval signal at c-terminal
Resident ER membrane protein
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Many proteins are selectively retained in the compartments in
which they function
Aggregation of proteins that function in the same compartment- called kin recognition
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The golgi apparatus consists of an ordered series of compartments.
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Two major classes of N-linked oligosaccharide
complex
High mannoseNo new sugar added in Golgi
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Oligosaccharide processing in ER and Golgi
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High specificendoglycosidase
Can distinguish betweenthese two type
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Proteoglycans are assembled in the Golgi Apparatus
O-linked glycosylation
Proteoglycans are secreted or anchored to plasma membrane
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What is the purpose of N-glycosylation?
1. N-linked is prevalent in all eukaryotes, but absent in procaryotes.2. limited flexibility.3. Recognition4. Regulation of development5. Protective coat unit6. Cell-cell adhesion
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N-linked
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The golgi cisternae are organized as a series of processing compartments
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Functional compartmentalization
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Matrix proteins form a dynamic scaffold that helps organize the
apparatus
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Transport from trans golgi network to lysosomes
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Lysosome are the principal sitesof intracellular digestion
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Lysosomes are heterogeneous(morphology)
The diversity reflects the wide range of digestive function.
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Plant and fungal vacuoles are remarkably verstile lysosomes
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Controlling size
Vacuole function:Storage, degrade, cell size, turgr pressure, homostatic device
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Three pathways to degradation in lysosomes.
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M6P receptor recognizes lysosomal proteins in the Trans Golgi network (TGN)
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The M6P receptor shuttles between specific membranes
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Transport into the cell from the plasma membrane:
endocytosis
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Transport from the trans golgi network to the cell exterior:
exocytosis
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Chapter 13 practice
1. The endocytic and biosynthetic-secretory and retrieval pathway.2. Utilization of different coats in vesicular traffic.3. The role of SNAREs and SNAREs-interacting proteins in vesicle
transport .4. The traffic role of COPI &COPII coat shuttling between Golgi and ER. 5. What is KDEL signal and its role in retrieval pathway.6. How to distinguish between the high-mannose oligosaccharide and
complex oligosaccharide.7. Write the full name for Glc NAc, NANA(sialic acid).8. The functional compartmentization of Golgi apparatus.9. Three pathways to degradation in lysosomes.10. How to transport lysosomal enzyme to lysosome from ER.11. Possible fates for endocytosed transmembrane receptor proteins. 12. The receptor-mediated endocytosis of LDL.13. Three best-understanding pathways of protein sorting in trans Golgi.