1. Master class Biomolecular Sciences Part 1: Molecular Cell Biology. 4 September 2007 Paul van Bergen en Henegouwen

2. Master class Biomolecular Sciences Part 1: Molecular Cell Biology. Sept-Nov 08 04-09-08: Paul van Bergen en Henegouwen. Mechanisms of clathrin-independent endocytosis 11-09-08: Willem Stoorvogel. Endocytosis and MHC classII 18-09-08: X-track symposium 25-09-08: Ger Strous. Ubiquitination and endocytosis 02-10-08: Catherine Rabouille (coordinator). Classical and unconventional secretion 09-10-08:Madelon Maurice. Wnt pathway and membrane traffic 16-10-08: ABC-seminar I&I +IB Aio dag 23-10-08: Fulvio Reggiori. Autophagy 30-10-08: Bernd Helms.Lipid droplet biogenesis 06-11-08: Exam

3. Classical secretion unconventional Non clathrin mediated endocytosis

4. Before I start, few statements: http://www.bio.uu.nl/BiomolecularSciences/ - The course is compulsary. If you have no choice of missing ONE class, contact the teacher and the coordinator by email (on website) - Each lecture will be posted on the website few days after each class - Article presentation. Each of you will study one article per block in a group of 2-3- 4 persons. Prepare the article as well as possible while contacting the teacherat least one week in advance. When you meet, the preparation should be done and you come for questions and for sorting out details. - Read the article : Download the article from the web for the colour pictures before coming to the course using the guidelines, your brain and your knowledge. The more you have thought of it, the better the discussion. - Exam. There will be 7 questions related to the 7 articles presented, hence the above. The more you participate, the easiest. - Switch off your mobile. No ringing, no SMS.

5. Mechanisms of clathrin- independent internalization Paul van Bergen en Henegouwen

6. Endocytosis: what’s the problem? bendingfission

7. Five ways to bend a membrane

8. Clathrin-dependent internalization

9. Clathrin-coated and vesicles: Ultrathin section of plastic-embedded material

10. Clathrin-coated pits and vesicles by EM Replica of freeze-dried plasma membranes

11. Clathrin-coated vesicles: Replica of freeze-dried plasma membranes

12. Clathrin: Heavy- and light chains

13. Clathrin makes a network

14. How clathrin is bending

15. Adaptor Protein-2 complex, AP-2     Ear Hinge Trunk 100 kDa 50 kDa 17 kDa  -adaptin  -adaptin

16. There are different AP-complexes on different sites in the cell Golgi-complex Plasma membrane Golgi-complex            AP-1 AP-2 AP-3

17. Protein/protein interactions control CCV formation     Receptor Yxx  Eps15 DPF Receptor LL L  x  D/E Clathrin AP-2 I EHs II COIL III DPF/UIM Epsin NPF Lipids DPW PIP 2

18. The netwerk forms a bal: Electron tomography of a coated vesicle ©John Wiley &Sons,Inc Adaptor protein (AP2) complex  chain  chain  adaptin  adaptin Clathrin triskelion Smith et al. (1998) EMBO J. 17, 4943

19. Overview of clathrin structures Marsh and McMahon (1999) Science 285, 215-220

20. From bending to fission

21. Dynamin, a GTPase involved in fission PIP 2 Amphiphysin

22. Dynamin is forming a ring around the neck

23. Effect of pure dynamin on lipid vesicles

24. PIPK Membrane curvature generation AP2 Clathrin Eps15 Intersectin Amphiphysin Epsin Dynamin Endophilin Moleculen involved in the whole process

25. Mogelijke rol van actine filamenten

26. PIPK Membrane curvature generation AP2 Clathrin Eps15 Intersectin Amphiphysin Epsin Dynamin Endophilin Actin Synaptojanin Auxilin Hsc70 Moleculen betrokken bij CCV vorming

27. Overview of all interactions

28. EGFR internalization via a non-coated pathway!

29. Is the alternative route visible?

30. Caveolae

31. Caveolin

32. Caveolar internalization starting from cholesterol-rich domains

33. Trafficking of Caveolae

34. Internalization of large items: Phagocytosis

35. Overview of different internalization routes

36. Properties of different CI-routes

37. Classification of internalization routes

38. Questions What does the bending? What does the fission?

39. Interleukin-2 receptor complex

40. IL-2R internalization is Clathrin-independent clathrin-inhibition by ΔEps15

41. IL-2R internalization is Dynamin-dependent Dynamin-inhibition by Dyn-K44 TfR wt K44A

42. IL-2R is present in the Detergent Resistant Membrane (DRM) fraction

43. Overview of different internalization routes