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Cs28: TMV: Sub-assembly, recapitulation, and control

Cs28: TMV: Sub-assembly, recapitulation, and control. Biochemistry 655 20 March 2011. J.D. Watson’s Post-doc. Goals:. Celebrate the work of Aaron Klug: Pragmatic Eclectic par excellence Understand phase diagrams! Sub-assembly: The TMV Disk

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Cs28: TMV: Sub-assembly, recapitulation, and control

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  1. Cs28: TMV:Sub-assembly, recapitulation, and control Biochemistry 655 20 March 2011

  2. J.D. Watson’s Post-doc

  3. Goals: • Celebrate the work of Aaron Klug: Pragmatic Eclectic par excellence • Understand phase diagrams! • Sub-assembly: The TMV Disk • Review the role of carboxylate residues in creating high-affinity proton binding • Review cooperativity and allostery! The disk-to-lockwasher transition • Understand how the concept of DGr works in the disk-to-lockwasher transition • Using rate measurements to distinguish initiation from elongation • Saturation behavior • Protein concentration dependence and ”molecularity” • RNA concentration-dependence and “specificity” • Understand the origins of RNA sequencing and footprinting!! • Recycling a good mechanism: recapitulation

  4. Eclectic Pragmatism: Klug’s trademark * ** Aaron Klug Nobel Prize Chemistry 1982 * * ** * * *

  5. Out of the dustbin…onto the front page

  6. Experimental parameters & Changes of “state” Electron Microscopy Phase Diagram Analysis

  7. DGr Residual structural packing free energy Disk (favored at high pH) Protein helix (favored only at low pH)

  8. Poly A TMV [RNA] dependence [Protein] dependence Nucleation and Growth

  9. 5’ 3’ TMV Recognition sequence

  10. Jaws (1975, Stephen Spielberg)

  11. Stripped Assembled Assembling is not the same as stripping!

  12. Assembly rates differ

  13. “Recycling” what is good about a mechanism

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