Authors: Nobuhisa Umeki¹; Tsuyoshi Yoshizawa¹; Yasunobu Sugimoto²; Toshiaki Mitsui³; Katsuzo Wakabayashi²; Shinsaku Maruta¹

Source: The Journal of Biochemistry, Volume 136, Number 6, 01 December 2004 , pp. 839-846(8)

Publisher: Oxford University Press

Abstract:

The region containing reactive cysteines, Cys 707 (SH1)–Cys 697 (SH2), of skeletal muscle myosin is thought to play a key role in the conformational changes of the myosin head during force generation coupled to ATP hydrolysis. In the present study, we synthesized a photochromic crosslinker, 4,4-azobenzene-dimaleimide (ABDM), that undergoes reversible cis-trans isomerization upon ultra violet (UV) and visible (VIS) light irradiation resulting in a change in the crosslinking length from 5 to 17 Å. The reactive cysteines, SH1 and SH2, of myosin subfragment 1 (S1) were crosslinked with ABDM, yielding an ABDM-S1 complex. The changes in absorbance induced by UV/VIS light irradiation of the complex were similar to those of free ABDM indicating that the incorporation of ABDM at the SH1 and SH2 sites did not disrupt the isomerization of crosslinked ABDM. Small-angle synchrotron X-ray scattering analysis of the ABDM-S1 complex in solution suggested that the localized conformational changes resulting from the cis to trans isomerization on ABDM crosslinking of SH1 and SH2 induced a small but significant swing in the lever arm portion of S1 in the opposite direction from that induced by ATP.

Document Type: Research article

DOI: http://dx.doi.org/10.1093/jb/mvh194

Affiliations: 1: Division of Bioengineering, Graduate School of Engineering, Soka University, Hachioji, Tokyo 192-8577; 2: Division of Biophysical Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka 560-8531 3: Group of Plant and Microbial Genome Controls, Graduate School of Science and Technology, Niigata University, Niigata 950-21; and

Publication date: 2004-12-01

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