Cosolvent-Induced Aggregation Inhibits Myosin ATPase Activity by Stabilizing the Predominant Transition Intermediate

Y. Michael Peyser, Shirley Shaya, Katalin Ajtai, Thomas P Burghardt, Andras Muhlrad

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

High concentration of the cosolvent poly(ethylene glycol) (PEG) induces reversible aggregation of skeletal myosin subfragment 1 (S1) and inhibition of its Mg-ATPase activity [Highsmith et al. (1998) Biophys. J. 74, 1465-1472]. In the present work the effect of aggregation on the various steps of the ATPase cycle was studied. The isomerization and hydrolysis steps of the cycle were not affected by S1 aggregation since the formation of the "trapped" S1·MgADP·phosphate analogue complexes, which mimic the prehydrolysis M*ATP and posthydrolysis M**ADP·P i transition states, proceeded without any hindrance. Similar conclusions could be reached from the chemical modification of Lys-83 and Cys-707 in the presence of MgATP and MgATPγS, which indicated that the most populated intermediate of the cycle in solubilized and aggregated S1 is M**ADP·Pi. The dissociation of the trapped S1·MgADP· phosphate analogue complexes resembling the M**ADP·Pi state was strongly inhibited by PEG-6000, showing that the transition from this intermediate is prevented by the aggregation. This step is presumably inhibited because the coupled swinging of the lever arm from the closed to the open position is constrained by the close packing of aggregated S1.

Original languageEnglish (US)
Pages (from-to)12669-12675
Number of pages7
JournalBiochemistry
Volume42
Issue number43
DOIs
StatePublished - Nov 4 2003

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Myosins
Adenosine Triphosphatases
Agglomeration
Adenosine Triphosphate
Myosin Subfragments
Ethylene Glycol
Polyethylene glycols
Hydrolysis
Phosphates
Chemical modification
Isomerization
Polyethylene Glycol 6000

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cosolvent-Induced Aggregation Inhibits Myosin ATPase Activity by Stabilizing the Predominant Transition Intermediate. / Peyser, Y. Michael; Shaya, Shirley; Ajtai, Katalin; Burghardt, Thomas P; Muhlrad, Andras.

In: Biochemistry, Vol. 42, No. 43, 04.11.2003, p. 12669-12675.

Research output: Contribution to journalArticle

Peyser, Y. Michael ; Shaya, Shirley ; Ajtai, Katalin ; Burghardt, Thomas P ; Muhlrad, Andras. / Cosolvent-Induced Aggregation Inhibits Myosin ATPase Activity by Stabilizing the Predominant Transition Intermediate. In: Biochemistry. 2003 ; Vol. 42, No. 43. pp. 12669-12675.
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