Raman difference spectroscopic studies of the myosin S1·MgADP·Vanadate complex

Hua Deng, Jianghua Wang, Robert Callender, Jean C. Grammer, Ralph G. Yount

Research output: Contribution to journalArticle

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Abstract

The Raman spectra of the nonbridging V-O bonds in the myosin S1·MgADP·Vi complex, often believed to be a transition-state analogue for the phosphotransfer reaction catalyzed by myosin, and in a vanadate solution model compound have been obtained using Raman difference spectroscopic techniques. A symmetric/asymmetric pair of modes at 870 cm-1 is found for vanadate in solution while three bands are found in the myosin S1·MgADP·Vi complex at 870, 844, and 829 cm-1. Using empirical relationships that relate bond order/bond lengths to stretch frequencies, the bond order and bond length of the three nonbridging V-O bonds of vanadate in solution were determined to be 1.43 vu (±0.04 vu) and 1.669 Å (±0.004 Å) respectively. The average bond order and bond length of the nonbridging V-O bonds in the S1·MgADP·Vi complex were determined to be 1.38 vu and 1,683 Å. A normal- mode analysis suggests that the VO32- moiety approaches a planar conformation in the enzymic complex. Ab initio calculations show that a water molecule at the S1 ATPase binding site, in line with the apical O-V bond in the ADP-Vi moiety and believed to be the attacking nucleophile in the phosphotransfer reaction, can account well for the changes in frequencies of vanadate when it binds to the protein by forming a moderately strong V- O(H2) bond. Hence, an important role determining the ATPase activity at the active site of myosin appears to be a strategic positioning of this in-line water molecule. Assuming that the distortions that vanadate undergoes upon forming the S1·MgADP·Vi complex are analogous to the changes of the γ- phosphate of ATP in the transition state of the myosin-catalyzed hydrolysis, our results suggest that this reaction proceeds close to a concerted (SN2- like) process.

Original languageEnglish (US)
Pages (from-to)10972-10979
Number of pages8
JournalBiochemistry
Volume37
Issue number31
DOIs
StatePublished - Aug 4 1998

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Vanadates
Myosins
Bond length
Adenosine Triphosphatases
Water piping systems
Nucleophiles
Molecules
Water
Chemical bonds
Adenosine Diphosphate
Conformations
Raman scattering
Hydrolysis
Catalytic Domain
Adenosine Triphosphate
Phosphates
Binding Sites
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Raman difference spectroscopic studies of the myosin S1·MgADP·Vanadate complex. / Deng, Hua; Wang, Jianghua; Callender, Robert; Grammer, Jean C.; Yount, Ralph G.

In: Biochemistry, Vol. 37, No. 31, 04.08.1998, p. 10972-10979.

Research output: Contribution to journalArticle

Deng, Hua ; Wang, Jianghua ; Callender, Robert ; Grammer, Jean C. ; Yount, Ralph G. / Raman difference spectroscopic studies of the myosin S1·MgADP·Vanadate complex. In: Biochemistry. 1998 ; Vol. 37, No. 31. pp. 10972-10979.
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title = "Raman difference spectroscopic studies of the myosin S1·MgADP·Vanadate complex",
abstract = "The Raman spectra of the nonbridging V-O bonds in the myosin S1·MgADP·Vi complex, often believed to be a transition-state analogue for the phosphotransfer reaction catalyzed by myosin, and in a vanadate solution model compound have been obtained using Raman difference spectroscopic techniques. A symmetric/asymmetric pair of modes at 870 cm-1 is found for vanadate in solution while three bands are found in the myosin S1·MgADP·Vi complex at 870, 844, and 829 cm-1. Using empirical relationships that relate bond order/bond lengths to stretch frequencies, the bond order and bond length of the three nonbridging V-O bonds of vanadate in solution were determined to be 1.43 vu (±0.04 vu) and 1.669 {\AA} (±0.004 {\AA}) respectively. The average bond order and bond length of the nonbridging V-O bonds in the S1·MgADP·Vi complex were determined to be 1.38 vu and 1,683 {\AA}. A normal- mode analysis suggests that the VO32- moiety approaches a planar conformation in the enzymic complex. Ab initio calculations show that a water molecule at the S1 ATPase binding site, in line with the apical O-V bond in the ADP-Vi moiety and believed to be the attacking nucleophile in the phosphotransfer reaction, can account well for the changes in frequencies of vanadate when it binds to the protein by forming a moderately strong V- O(H2) bond. Hence, an important role determining the ATPase activity at the active site of myosin appears to be a strategic positioning of this in-line water molecule. Assuming that the distortions that vanadate undergoes upon forming the S1·MgADP·Vi complex are analogous to the changes of the γ- phosphate of ATP in the transition state of the myosin-catalyzed hydrolysis, our results suggest that this reaction proceeds close to a concerted (SN2- like) process.",
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AB - The Raman spectra of the nonbridging V-O bonds in the myosin S1·MgADP·Vi complex, often believed to be a transition-state analogue for the phosphotransfer reaction catalyzed by myosin, and in a vanadate solution model compound have been obtained using Raman difference spectroscopic techniques. A symmetric/asymmetric pair of modes at 870 cm-1 is found for vanadate in solution while three bands are found in the myosin S1·MgADP·Vi complex at 870, 844, and 829 cm-1. Using empirical relationships that relate bond order/bond lengths to stretch frequencies, the bond order and bond length of the three nonbridging V-O bonds of vanadate in solution were determined to be 1.43 vu (±0.04 vu) and 1.669 Å (±0.004 Å) respectively. The average bond order and bond length of the nonbridging V-O bonds in the S1·MgADP·Vi complex were determined to be 1.38 vu and 1,683 Å. A normal- mode analysis suggests that the VO32- moiety approaches a planar conformation in the enzymic complex. Ab initio calculations show that a water molecule at the S1 ATPase binding site, in line with the apical O-V bond in the ADP-Vi moiety and believed to be the attacking nucleophile in the phosphotransfer reaction, can account well for the changes in frequencies of vanadate when it binds to the protein by forming a moderately strong V- O(H2) bond. Hence, an important role determining the ATPase activity at the active site of myosin appears to be a strategic positioning of this in-line water molecule. Assuming that the distortions that vanadate undergoes upon forming the S1·MgADP·Vi complex are analogous to the changes of the γ- phosphate of ATP in the transition state of the myosin-catalyzed hydrolysis, our results suggest that this reaction proceeds close to a concerted (SN2- like) process.

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