P1249616421uXJdQ

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CONFORMATIONAL CHANGES AT THE NUCLEOTIDE-BINDING
SITE OF KINESIN-FAMILY MOTORS ED PATE WSU
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FORCE GENERATION IN MUSCLE IS PRODUCED BY THE
INTERACTION OF TWO PROTEINS ACTIN AND MYOSIN
MYOSIN
ACTIN
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MYOSIN KINESIN
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NUCLEOTIDE BINDING SITE
MYOSIN KINESIN
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COMPARISON OF NCD AND MYOSIN BINDING SITES
NCD
MYOSIN
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LOCATION OFKINESIN CYS-188
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VANT HOFF ANALYSIS SHOWS DH0 -100 KJ/MOL FOR
THE TRANSITION
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SUMMARY OF CONE ANGLE CHANGES ON BINDING TO
MICROTUBULES
ANALOG D ANGLE EPR
ncd
SSL-NANDP 44
SL-NANDP gt28
2'-SLADP 12
3'-SLADP gt12
2', 3'-SLADP gt16

kinesin
SSL-NANDP 35
SL-NANDP 32
2'-SLADP gt10
3'-SLADP 11
2', 3'-SLADP gt18
12
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12 Å
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POTENTIAL ENERGY COMPONENTS FROM BONDED
INTERACTIONS
Stretch
Flex
Torsion
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POTENTIAL ENERGY COMPONENTS FROM NON-BONDED
INTERACTIONS
Coulomb Interaction
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THE OPEN SWITCH 1 CONFORMATION IS STABLE
ADP
SWITCH 1
P-LOOP
P-LOOP
SWITCH 1
SWITCH 2
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THE CLOSED SWITCH 1 CONFORMATION IS STABLE
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INFLUENCE OF SWITCH 1 ON EPR PROBES
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ANGULAR DISTRIBUTION FOR SSL-NANDP IN THE OPEN
AND CLOSED CONFORMATIONS
-90
90
0
ANGLE
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COORDINATION AT THE OPEN NUCLEOTIDE SITE
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COORDINATION AT THE CLOSED NUCEOTIDE SITE
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MYOSIN KINESIN
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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.

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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