The kinetics of a two-state transition of myosin subfragment 1. A temperature-jump relaxation study.

Abstract:

:Temperature-jump measurements were carried out on myosin subfragment 1 (S1) labeled at Cys-707 with 5-(iodoacetamido)fluorescein (S1-AF). The relaxation was monitored by following the increase in the fluorescence intensity of the attached probe after a jump of 5.8 degrees C. A single relaxation process was observed over a range of final temperatures, and the relaxation time decreased from 16.69 ms at 15 degrees C to 3.91 ms at 27 degrees C. The relaxation results are interpreted in terms of a two-state transition: (S1-AF)L K+ in equilibrium with K- (S1-AF)H, and the observed single relaxation time (tau) equals l/(k(+) + k-). The individual first-order rate constants, k+ and k-, were calculated from tau and the equilibrium constant previously determined. The activation energy was 21.9 kcal/mol for the forward reaction and 9.3 kcal/mol for the reverse reaction, corresponding to an enthalpy value of 12.6 kcal/mol for the two-state transition. The results provide, for the first time, direct kinetic evidence of a two-state transition of S1 in the absence of bound nucleotide, and support a two-state model of unliganded myosin subfragment 1.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Lin SH,Cheung HC

doi

10.1016/0014-5793(92)80614-m

keywords:

subject

Has Abstract

pub_date

1992-06-15 00:00:00

pages

184-6

issue

2-3

eissn

0014-5793

issn

1873-3468

pii

0014-5793(92)80614-M

journal_volume

304

pub_type

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