ANAT 262 Lecture Notes - Lecture 21: Differential Centrifugation, Secretion, Golgi Cell

• School: McGill University
• Department: Anatomy & Cell Biology
• Course: ANAT 262
• Professor: Ursula Stochaj

Document Summary

Cargo starts in er golgi cell surface/endocytic pathway. What happens in golgi using in vitro systems. Differential centrifugation (short), followed by density gradient centrifugation (longer) Can also use to purify golgi instead, last step is density gradient centrifugation. Reproduce process occurring inside the cell, outside the cell. Uncharacterized cytosol is mix of many proteins in cytoplasm proteins replace w/the few proteins essential to process. Vs. using tagged protein to study process inside cell, very complicated. Break open cell homogenize er break down into vesicles (microsomes), still er differential centrifugation get rid of heavy (e. g. nuclei) . Density gradient centrifugation separate smooth & rough microsomes. Confirm cytoplasmic proteins synthesized at ribosomes, proteins in secretory pathway. Ribosome docking at er, proteins into er & fold. Detergent unfolds proteins, linearizes, charge negatively go towards positive electrode larger protein travel slower than smaller. Assay determining whether proteins translocate into er based on ses page.