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12 - Kuo 580.221 17 Oct 2006 Molecules Cells BME 580.221...

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Kuo, 580.221 17 Oct 2006 1 Molecules & Cells BME 580.221 Protein Secretion/Trafficking Lecture #12, 17 Oct 2006 Scot C. Kuo 17 Oct 2006 Kuo, 580.221 How to study protein trafficking 17 Oct 2006 Kuo, 580.221 Which membrane pathways leads to the outside of the cell? Endoplasmic reticulum and Golgi bodies Are these membranes like mitochondria? 17 Oct 2006 Kuo, 580.221 Smooth & rough ER 17 Oct 2006 Kuo, 580.221 Free vs Membrane Bound Ribosomes 17 Oct 2006 Kuo, 580.221 Expt: lumenal location of secretory proteins Tools : proteases, detergents, protein gels (polyacrylamide electrophoresis & SDS; Westerns) >Show Flash Animations< Microsomes are biochemically purified versions of endoplasmic reticulum
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Kuo, 580.221 17 Oct 2006 2 17 Oct 2006 Kuo, 580.221 Expt: Evidence for CoTranslational Insertion 17 Oct 2006 Kuo, 580.221 Crossing the ER membrane 17 Oct 2006 Kuo, 580.221 Translocation of soluble protein 17 Oct 2006 Kuo, 580.221 Mito. vs ER protein processing Yes -- many ?? Translation req’d for translocation SRP, then SRP receptor on ER ER no no Complete protein will translocate receptor on Mito Mito Add’l trafficking Add’l Protein Modifications Protein Signal Seq Binding 17 Oct 2006 Kuo, 580.221 Translocation of soluble protein (Ab?) 17 Oct 2006 Kuo, 580.221 Integration of transmembrane protein
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Kuo, 580.221 17 Oct 2006 3 17 Oct 2006 Kuo, 580.221 Integration of double-pass transmembrane protein 17 Oct 2006 Kuo, 580.221 Questions for Ab secretion How to assemble? 4 polypeptides multiple disulfide bonds between peptides addition of carbohydrate (sugars) to peptide (common feature of secreted proteins; why?) How to move from ER to outside cell?
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