Intradomain distances in the regulatory domain of the myosin head in prepower and postpower stroke states

Fluorescence energy transfer

Thomas Palm, Ken Sale, Louise Brown, Huichun Li, Brett Hambly, Peter G. Fajer*

*Corresponding author for this work

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The relative movement of the catalytic and regulatory domains of the myosin head (S1) is likely to be the force generating conformational change in the energy transduction of muscle [Rayment, I., Holden, H. M., Whittaker, M., Yohn, C. B., Lorenz, M., Holmes, K. C., and Milligan, R. A. (1993) Science 261, 58-65]. To test this model we have measured, using frequency- modulated FRET, three distances between the catalytic domain and regulatory domains and within the regulatory domain of myosin. The donor/acceptor pairs included MHC cys707 and ELC cys177; ELC cys177 and RLC cys154; and ELC cys177 and gizzard RLC cys108. The IAEDANS (donor) or acceptor (DABMI or IAF) labeled light chains (ELC and RLC) were exchanged into monomeric myosin and the distances were measured in the putative prepower stroke states (in the presence of MgATP or ADP/AlF664/-) and the postpower stroke states (ADP and the absence of nucleotides). For each of the three distances, the donor/acceptor pairs were reversed to minimize uncertainty in the distance measured, arising from probe orientational factors. The distances obtained from FRET were in close agreement with the distances in the crystal structure. Importantly, none of the measured distances varied by more than 2 Å, putting a strong constraint on the extent of conformational changes within S1. The maximum axial movement of the distal part of myosin head was modeled using FRET distance changes within the myosin head reported here and previously. These models revealed an upper bound of 85 Å for a swing of the regulatory domain with respect to the catalytic domain during the power stroke. Additionally, an upper bound of 22 Å could be contributed to the power stroke by a reorientation of RLC with respect to the ELC during the power stroke.

Original languageEnglish
Pages (from-to)13026-13034
Number of pages9
JournalBiochemistry
Volume38
Issue number40
DOIs
Publication statusPublished - 5 Oct 1999
Externally publishedYes

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