Open AccessCoupling of nonpolymerizable monomeric actin to the F-actin binding region of the myosin head.
Author(s) -
Nadir Bettache,
Raoul Bertrand,
Ridha Kassab
Publication year - 1989
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.86.16.6028
Subject(s) - maleimide , myosin , actin , chemistry , covalent bond , monomer , intramolecular force , myosin head , tropomyosin , pyrophosphate , actina , biochemistry , stereochemistry , myosin light chain kinase , biophysics , polymer , cytoskeleton , polymer chemistry , biology , organic chemistry , enzyme , cell
Polymerizations of skeletal G-actin induced by salt and myosin subfragment 1 (S-1) were suppressed by reaction of G-actin with m-maleimidobenzoyl-N-hydroxysuccinimide ester. The G-actin derivative, containing few intramolecular crosslinks and a free maleimide group, was covalently coupled in solution to the S-1 heavy chain. The resulting complex could no longer bind to F-actin. The SH-1 and SH-2 thiols of S-1 were not involved in the complexation and the covalent link was shown to be exclusively on the 50-kDa segment of the S-1 heavy chain. The specific conjugation of the two proteins followed formation of a reversibly associated pyrophosphate-sensitive binary complex which was characterized by different approaches. Potentially, these complexes may be useful in developing the crystallography of actin-bound S-1.