Cross-linking of three heavy-chain domains of myosin adenosinetriphosphatase with a trifunctional alkylating reagent.
The chemotherapeutic alkylating reagent tris(2-chloroethyl)amine (TCEA) was used as a trifunctional cross-linking reagent with a cross-linking span of 5 A for myosin subfragment 1 (S-1). When S-1 was incubated with TCEA, all three domains of 20, 26, and 50 kDa in the S-1 heavy chain were cross-linked via the highly reactive sulfhydryl group SH1 (Cys-707) on the 20-kDa domain. The cross-linking was accelerated by nucleotides. The present observation is consistent with the proposal that SH1 is close to both the 26- and 50-kDa domains of S-1 and that movement within S-1 associated with the nucleotide binding occurs around SH1 as well as around another reactive thiol, SH2 & Wong, A. G. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 6392-6396; Hiratsuka, T. (1987) Biochemistry 26, 3168-3173].
Hiratsuka
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Department of Chemistry, Asahikawa Medical College, Hokkaido, Japan.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000477:Alkylating Agents / Q000494:pharmacology*; D000478:Alkylation; D000818:Animals; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D009588:Nitrogen Mustard Compounds / Q000494:pharmacology*; D009711:Nucleotides / Q000494:pharmacology; D011817:Rabbits; D014357:Trypsin
D000477:Alkylating Agents; D003432:Cross-Linking Reagents; D009588:Nitrogen Mustard Compounds; D009711:Nucleotides; C009802:2,2',2''-trichlorotriethylamine; D014357:Trypsin; D000251:Adenosine Triphosphatases