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CONFORMATIONAL CHANGES AT THE NUCLEOTIDE-BINDING SITE OF KINESIN-FAMILY MOTORS ED PATE WSU

CONFORMATIONAL CHANGES AT THE NUCLEOTIDE-BINDING SITE OF KINESIN-FAMILY MOTORS ED PATE WSU. FORCE GENERATION IN MUSCLE IS PRODUCED BY THE INTERACTION OF TWO PROTEINS: ACTIN AND MYOSIN. MYOSIN. ACTIN. MYOSIN KINESIN. NUCLEOTIDE BINDING SITE. MYOSIN KINESIN.

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CONFORMATIONAL CHANGES AT THE NUCLEOTIDE-BINDING SITE OF KINESIN-FAMILY MOTORS ED PATE WSU

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  1. CONFORMATIONAL CHANGES AT THE NUCLEOTIDE-BINDING SITE OF KINESIN-FAMILY MOTORS ED PATE WSU

  2. FORCE GENERATION IN MUSCLE IS PRODUCED BY THE INTERACTION OF TWO PROTEINS: ACTIN AND MYOSIN MYOSIN ACTIN

  3. MYOSIN KINESIN

  4. NUCLEOTIDE BINDING SITE MYOSIN KINESIN

  5. COMPARISON OF NCD AND MYOSIN BINDING SITES MYOSIN NCD

  6. LOCATION OFKINESIN CYS-188

  7. VAN’T HOFF ANALYSIS SHOWS DH0 = -100 KJ/MOL FOR THE TRANSITION

  8. SUMMARY OF CONE ANGLE CHANGES ON BINDING TO MICROTUBULES

  9. CONSTRUCTION OF NCD WITH SWITCH 1 CLOSED ADP P-LOOP NCD SWITCH 1 SWITCH 2

  10. CONSTRUCTION OF NCD WITH SWITCH 1 CLOSED MYOSIN SWITCH 1 P-LOOP NCD SWITCH 1 SWITCH 2

  11. CONSTRUCTION OF NCD WITH SWITCH 1 CLOSED MODIFIED NCD SWITCH 1 ADP P-LOOP NCD SWITCH 1 SWITCH 2 12 Å

  12. POTENTIAL ENERGY COMPONENTS FROM BONDED INTERACTIONS Stretch Flex Torsion

  13. POTENTIAL ENERGY COMPONENTS FROM NON-BONDED INTERACTIONS Coulomb Interaction

  14. THE OPEN SWITCH 1 CONFORMATION IS STABLE ADP SWITCH 1 P-LOOP P-LOOP SWITCH 1 SWITCH 2

  15. SWITCH 1 SWITCH 1 P-LOOP P-LOOP SWITCH 2 THE CLOSED SWITCH 1 CONFORMATION IS STABLE

  16. MOLECULAR DYNAMICS RESULTS • Open structure (x-ray) stable at 300 K (ADP, ATP, APO) • Closed structure (model) stable at 300 K (ADP, ATP, APO) Energy barrier Open and closed structures Are local minima on a global free energy surface. Test with high temperature Dynamics. G Closed Open

  17. INFLUENCE OF SWITCH 1 ON EPR PROBES

  18. ANGULAR DISTRIBUTION FOR SSL-NANDP IN THE OPEN AND CLOSED CONFORMATIONS -90 0 90 ANGLE

  19. COORDINATION AT THE OPEN NUCLEOTIDE SITE

  20. COORDINATION AT THE CLOSED NUCEOTIDE SITE

  21. MYOSIN KINESIN

  22. CONCLUSIONS Comparison with myosin suggests a new structural state of kinesin-family motors involving a closing of the nucleotide site. 2. Spectroscopic data show the nucleotide site closes when kinesin binds to microtubules. 3. MD shows the open and closed binding-site conformations of kinesin-family motors explain the changes in mobility of nucleotide analog EPR probes upon binding to microtubules. MD shows the closed state is essential for nucleotide hydrolysis and force production.

  23. COLLABORATORS ROGER COOKE, UCSF RALPH YOUNT, WSU Nariman Naber Xiaoru Chen Marija Matuska Jean Grammer Kathy Franks-Skiba PETER KOLLMAN, UCSF RON VALE, UCSF Todd Minehardt Sarah Rice ROBERTO CAR, PRINCETON DAVID ADCOCK, LONE STAR BIOTECH

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