ANAT 262 Lecture Notes - Lecture 14: Collagen, Secretion, Lipid Raft

• School: McGill University
• Department: Anatomy & Cell Biology
• Course: ANAT 262
• Professor: John Presley

Document Summary

Proteins are modified in all types of way in cisternae. In vitro system: purified golgi from two sources: (1) transferase mutant and virus infected (g-protein) and (2) transferase present, non-infected (no g protein). G protein would have to be sorted in vesicles, leaving cisternae for the other golgi. If glcnac was present on g protein, it suggests that transferase and g protein were found in same compartment. In vitro assay led to discovery of copi. Addition of non-hydrolyzable gtp analog resulted in accumulation of vesicles. Nsf: n-methyl-maleimide (nem) inactivates most protein, and block in vitro assay at low concentrations. It suggest that only one protein was targeted. When treated, vesicles were accumulated but not coated, suggesting that nsf is required for fusion. Treated cytosol with nem, and tried to rescue cytosol by monitoring glcnac incorporation. Components required for sorting/budding of some cargoes found (copi) Cytosolic proteins required for fusion found (led eventually to the discovery of.