6 - Intercompartmental Transport

6 - Intercompartmental Transport - Lecture 6:...

Info iconThis preview shows pages 1–9. Sign up to view the full content.

View Full Document Right Arrow Icon
Lecture 6: Intercompartmental protein transport technical concepts: EM established the early basic principles Reconstitution systems to identify components and mechanisms Genetic approaches to identify mutants Microscopy of living cells with activatable/inactivatable tracers
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
How we know what we know about intercompartmental protein transport 1. Electron microscop y establishe d the basic ideas G. Palade Nobel Lecture, Science Vol 189 p 347, 1975
Background image of page 2
Key concepts to emerge from Palade 1. Newly-synthesized Proteins pass through a series of membrane compartments on their way to cell surface 2. Transfer of proteins between these compartments is mediated by vesicles 3. Vesicular transport intermediates are characterized by spiky coats Amazingly, all these things are true!
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Subsequent understanding of mechanisms of intercompartmental transport have been driven largely by a combination of genetics and biochemistry
Background image of page 4
The genetics: Secretion is essential. How can you get mutations that block an essential process, but nevertheless the cells can proliferate and be sustained in culture? Concept of conditional lethality. Folded @ 23 Unfolded @ 37 Often degrade d
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
The genetics: Identification of mutants in Saccharomyces cerevisiae that are unable to secrete proteins @ 37°C, and as a result become dense Novick et al 1980 Cell 21:205-15 This screen yielded 23 Sec genes. Most Sec proteins you encounter come from this screen
Background image of page 6
Normal at permissive Vesicles pile up at NPT
Background image of page 7

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Wild type alleles of temperature- sensitive sec genes can be cloned by complementation sec18 mutant at 37°C: can’t grow introduce library of genomic DNA fragments, ~10 kb/plasmid vast majority
Background image of page 8
Image of page 9
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 01/12/2012 for the course BI 9 taught by Professor Aravin during the Spring '10 term at Caltech.

Page1 / 37

6 - Intercompartmental Transport - Lecture 6:...

This preview shows document pages 1 - 9. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online