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Master course Molecular & Cellular Life Sciences Part 1: Intracellular Membrane Processes October 8, 2009 Paul van Bergen en Henegouwen
Mechanisms of clathrin-independent internalization Paul van Bergen en Henegouwen
Different endocytic portals
Endocytosis: whats the problem? bendingfission
Five ways to bend a membrane
Clathrin-coated and vesicles: Ultrathin section of plastic-embedded material
Clathrin-coated pits and vesicles by EM Replica of freeze-dried plasma membranes
Clathrin-coated vesicles: Replica of freeze-dried plasma membranes
Clathrin: Heavy- and light chains
Clathrin makes a network
How clathrin is bending?
AP-2 Adaptor Protein-2 complex a b2b2 m2m2 s2s2 Ear Hinge Trunk 100 kDa 50 kDa 17 kDa a-adaptin b-adaptin
There are different AP-complexes on different sites in the cell Golgi-complex Plasma membrane Golgi-complex ab2b2 m2m2 s2s2 g b1b1 m1m1 s1s1 d b3b3 m3m3 s3s3 AP-1 AP-2 AP-3
Protein/protein interactions control CCV formation a b2 m2 s2 Receptor Yxxf Eps15 DPF Receptor LL L fxf D/E Clathrin AP-2 I EHs II COIL III DPF/UIM Epsin NPF Lipids DPW PIP 2
Epsin induces bending of liposomes
The netwerk forms a ball Electron tomography of a coated vesicle Adaptor protein (AP2) complex s chainm chain b adaptina adaptin Clathrin triskelion Smith et al. (1998) EMBO J. 17, 4943
Overview of clathrin structures Marsh and McMahon (1999) Science 285, 215-220
From bending to fission
Dynamin, a GTPase involved in fission PIP 2 Amphiphysin
Dynamin is forming a ring around the neck
Effect of pure dynamin on lipid vesicles
PIPK Membrane curvature generation AP2 Clathrin Eps15 Intersectin Amphiphysin Epsin Dynamin Endophilin Moleculen involved in the whole process
Possible role for actin filaments
PIPK Membrane curvature generation AP2 Clathrin Eps15 Intersectin Amphiphysin Epsin Dynamin Endophilin Actin Synaptojanin Auxilin Hsc70 Moleculen involved in the whole process
Overview of all interactions
EGFR internalization via a non-coated pathway!
Is the alternative route visible?
Caveolar internalization starting from cholesterol-rich domains
Trafficking of Caveolae
Phagocytosis: Internalization of large items
Overview of different internalization routes
Properties of different CI-routes
Classification of internalization routes
Questions What does the bending? What does the fission?
Interleukin-2 receptor complex
IL-2R internalization is Clathrin-independent clathrin-inhibition by ΔEps15
IL-2R internalization is Dynamin-dependent Dynamin-inhibition by Dyn-K44 TfR wt K44A
IL-2R is present in the Detergent Resistant Membrane (DRM) fraction
Master class Biomolecular Sciences Part 1: Molecular Cell Biology. 4 September 2007 Paul van Bergen en Henegouwen.
Cytoplasmic Membrane Systems: Structure, Function, and
Vesicle Formation at the Plasma Membrane and Trans-Golgi Network: The Same but Different M. A. McNiven, et al., Science 313, 1591 (2006) Department of.
Lecture 6 - Intracellular compartments and transport I.
Binding Studies on Trafficking Proteins Using Microcalorimetry
Dynamin - Monique Krüger - e4b07fbe88868c13%2F %2FBDLPxtal_map_filament.png&imgrefurl=http%3A%2F%2Fwww.thelowlab.org%2Fr
The ER and Golgi Modify & Sort Proteins
Molecular mechanism of vesicular transport Vesicular pathway--Budding and fusion 1.Budding— how are vesicles formed? How can cargo proteins be selectively.
Lecture 3 Vesicular Trafficking -Cops and Clathrins -Arfs, Rabs, Sars -Snares.
1.Phosphoinositide metabolism in synaptic vesicle recycling 2.Techniques in synaptic vesicle recycling.
Membranes in Biology Rob Phillips California Institute of Technology (McMahon)
Copyright © 2005 Pearson Prentice Hall, Inc. Intracellular Compartments and Transport Membrane Enclosed Organelles Protein Sorting Vesicular Transport.
Discovery of New Cargo Proteins that Enter Cells through Clathrin-Independent Endocytosis Craig A. Eyster, Jason D. Higginson, Robert Huebner, Natalie.
Most proteins in the ER are glycosylated Proteins in cytosol are rarely glycosylated original precursor oligosaccharide added to most proteins in the ER.
Endocytosis and Lysosome Formation The Endolytic Pathway.
INTD5000, Lectures C7 & C8 Professor Eileen M. Lafer Tel#: Office: Room 415B THE SECRETORY AND ENDOCYTIC PATHWAYS.
Introduction Question How are the high-molecular- weight yolk （卵黄） proteins able to enter the oocyte( 卵母细胞 )?
Endocytosis & Exocytosis By : Matt Sargeant. What is Endocytosis and Exocytosis? Endocytosis is a process by which cells absorb molecules (such as proteins)
Summary 1.Rough ER and smooth ER; 2.Signal hypothesis, translocation into ER; 3.Single-span and multi-span membrane proteins; 4.Glycosylation; 5.Protein.
SNARE complex and Ca 2+ sensor Endocytic invagination 1.Bar-domain containing protein such as amphiphysin which inducing membrane curvature 2.Clathrin,
Last Class: 1. transportation at the plasma membrane A. carrier protein, B. channel protein 2. intracellular compartments A. nucleus, B. Endoplasmic Reticulum.
Rust et al 2004Troels Scheel Single molecule biophysics Assembly of endocytic machinery around individual influenza viruses during viral entry Influenza.
Paul Cullen’s Lectures for BIO402/502 Office hours: Friday 2PM or by appointment Test will be on October 29th 7PM-9PM Short answer,
Vesicular transport Dr. med. habil. Kőhidai László Assoc. Professor Depertment of Genetics, Cell- and Immunoibiology Semmelwesi University Semmelwesi University2008.
Integral Proteins Some transmembrane transport proteins are porins that form aqueous channels through membranes Some integral proteins anchored to the.
Vesicular transport Dr. med. habil. Kőhidai László Assoc. Professor Depertment of Genetics, Cell- and Immunoibiology Semmelwesi University Semmelwesi University.
A Closer Look at Cell Membranes Aim: How do large particles enter and exit cells? Do Now: Name some molecules/materials that enter and exit the cell.
Previously Bio308 Hypotheses for molecular basis of bipolar disorder Suggest problem lies in protein targeting How are proteins targeted and delivered?
Lecture 7 - Intracellular compartments and transport II
Vesicular Transport III. N-linked glycosylation because sugar is added to N of asparagine. original precursor oligosaccharide added to most proteins in.
Receptor-Mediated Endocytosis As Presented By Michael Chan.
Vesicular Transport Pages The general flow of membrane enclosed material within cells.
Protein trafficking between membranes By Graham Warren & Ira Mellman
Dynamic networks & clathrin-mediated endocytosis Gerrit Praefcke (now at Cologne) Marijn Ford (now at UC Davis) Eva Schmid (LMB Cambridge)
Fractionation of organelles and membrane vesicles using OptiPrep™
Vesicular Traffic II. Endocytic and secretory pathways red = secretory green = endocytic blue = recycling.
Membrane Rafts ‘70s ‘40s‘50s. Membrane Microdomains Raft is a specific type of microdomain – sphingolipid/cholesterol rich region “Separation of discrete.
Cellular Structures and Organelles
The Nucleus Nuclear Organization Nuclear Envelope and Molecular Trafficking Nucleolus and rRNA Processing The nucleus is one of the main features that.
Receptor-Mediated Endocytosis Zhong huan zi Zhong huan zi.
Chapter 13 Intracellular Vesicular Traffic 張學偉 助理教授.
A Closer Look at Cell Membranes Chapter 5 Part 2.
Lecture 11 Vesicular Trafficking: Lysosomes;
Diffusion (more practice) Blue = water molecule (solvent) Red = salt molecule (solute) What will move where? How many? Do they just stop moving? Distilled.
Lecture 12: The secretory pathway
Chapter 5 of Yeagle Structure of Biological Membranes Non-lamellar phases Spontaneous curvature Actual curvature What is the physical basis of non-lamellar.
Cell Signaling and Migration Erich Lidstone April 29, 2009.
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