mcb110_alber_trafficking - Trafficking 1 MCB 110 - Fall...

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MCB 110 - Fall 2007- Alber Trafficking 1 4 MEMBRANE TRAFFICKING I Introduction: secretory pathway A. Protein Synthesis and sorting B. Methods to study cytomembranes II Endoplasmic Reticulum A. Smooth ER B. Rough ER C. Synthesis of proteins in membrane-bound ribosomes The signal hypothesis Synthesis of membrane proteins D. Glycosylation in the ER III Golgi IV Vesicle Transport A. COPII-coated Vesicles B. COPI-coated Vesoicles C. Clathrin-coated Vesicles V Lisosomes A. Phagocytosis B. Autophagy VI Endocytosis Suggested Reading: Lodish, Chapter 5, 5.3; Chapter 16, 16.1 to 16.3; Chapter 17 Alberts, Chapters 12 and 13
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MCB 110 - Fall 2007- Alber Trafficking 2 I Introduction to the Secretory Pathway The eukaryotic cell is filled with membranous organelles that form part of an integrated and dynamic system shuttling material across the cell
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MCB 110 - Fall 2007- Alber Trafficking 3 The biosynthetic or secretory pathway includes the synthesis of proteins in the ER, their modification in the ER and Golgi, and their transport to different destinations such as the plasma membrane, lysosomes, vacuoles, etc. In constitutive secretion materials are transported in a continual manner. In regulated secretion , materials are stored in secretory granules in the periphery of the cell and discarded in response to a particular stimulus (e.g. nerve cells, cells producing hormones or digestive enzymes). Materials are transported in vesicles that move along microtubules, powered by motor proteins. Sorting is facilitated by receptors localized in particular membranes.
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MCB 110 - Fall 2007- Alber Trafficking 4 Methods to Study Cytomembranes Visualization by electron microscopy Dynamic localization by autoradiography and pulse-chase
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MCB 110 - Fall 2007- Alber Trafficking 5
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MCB 110 - Fall 2007- Alber Trafficking 6 Use of GFP constructs Movement of proteins through the secretory pathway has been followed using a Green Flourescence Protein ( GFP ). Cells where infected with vesicular stomatitis virus (VSV) in which one of the genes (VSVG) is fused to GFP. Large amounts of VSVG protein are produced in the ER that move to the Golgi and then to the plasma membrane. The process can be seen as a wave of green fluorescence that can be synchronized using temperature mutants of VSVG than cannot leave the ER at high temperatures.
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MCB 110 - Fall 2007- Alber Trafficking 7 Subcellular fraction purification and characterization : When cells are homogenized the rough ER breaks up into small closed vesicles call microsomes .
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MCB 110 - Fall 2007- Alber Trafficking 8 Inmediately after their synthesis, secretory proteins are localized in the lumen of microsomes. If proteases are added to the
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This note was uploaded on 03/08/2010 for the course MCB 110 taught by Professor Alber,zhou,nogales during the Fall '07 term at University of California, Berkeley.

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mcb110_alber_trafficking - Trafficking 1 MCB 110 - Fall...

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