master course molecular & cellular life sciences part 1: intracellular membrane processes...
TRANSCRIPT
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Master course Molecular & Cellular Life Sciences
Part 1: Intracellular Membrane Processes
October 8, 2009
Paul van Bergen en Henegouwen
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Mechanisms of clathrin-independent
internalization
Paul van Bergen en Henegouwen
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Different endocytic portals
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Characteristics
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Endocytosis: what’s the problem?
bending fission
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Five ways to bend a membrane
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Clathrin-dependent internalization
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Clathrin-coated and vesicles:Ultrathin section of plastic-embedded material
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Clathrin-coated pits and vesicles by EMReplica of freeze-dried plasma membranes
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Clathrin-coated vesicles:Replica of freeze-dried plasma membranes
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Clathrin: Heavy- and light chains
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Clathrin makes a network
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How clathrin is bending?
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AP-2Adaptor Protein-2 complex
a b2m2
s2
EarHinge
Trunk
100 kDa
50 kDa
17 kDa
a-adaptin b-adaptin
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There are different AP-complexes on different sites in the cell
Golgi-complex Plasma membrane Golgi-complex
a b2m2
s2g b1m1
s1d b3
m3
s3
AP-1 AP-2 AP-3
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Protein/protein interactions control CCV formation
a b2m2s2
Receptor
Yxxf
Eps15
DPF
Receptor
LL
LfxfD/E
Clathrin
AP-2
I
EHs
II
COIL
III
DPF/UIM
Epsin
NPF
Lipids
DPW
PIP2
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Epsin induces bendingof liposomes
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The netwerk forms a ballElectron tomography of a coated vesicle
Adaptor protein(AP2) complex
s chain m chain
b adaptin
a adaptin
Clathrin triskelion
Smith et al. (1998) EMBO J. 17, 4943
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Overview of clathrin structures
Marsh and McMahon (1999) Science 285, 215-220
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From bending to fission
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Dynamin, a GTPase involved in fission
PIP2 Amphiphysin
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Dynamin is forming a ring around the neck
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Effect of pure dynamin on lipid vesicles
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PIPK
Membrane curvature generation
AP2ClathrinEps15
IntersectinAmphiphysinEpsin Dynamin
Endophilin
Moleculen involved in the whole process
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Possible role for actin filaments
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PIPK
Membrane curvature generation
AP2ClathrinEps15
IntersectinAmphiphysinEpsin Dynamin
Endophilin
Actin
SynaptojaninAuxilinHsc70
Moleculen involved in the whole process
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Overview of all interactions
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EGFR internalization via a non-coated pathway!
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Is the alternative route visible?
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Caveolae
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Caveolin
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Caveolar internalization starting from cholesterol-rich domains
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Trafficking of Caveolae
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Phagocytosis:Internalization of large items
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Overview of different internalization routes
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Properties of different CI-routes
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Classification of internalization routes
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Questions
What does the bending?What does the fission?
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Interleukin-2 receptor complex
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IL-2R internalization is Clathrin-independent clathrin-inhibition by ΔEps15
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IL-2R internalization is Dynamin-dependentDynamin-inhibition by Dyn-K44
TfR
wt
K44A
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IL-2R is present in the Detergent Resistant Membrane (DRM) fraction