Graphical evaluation of alkylation of myosin's SH1 and SH2: The N- phenylmaleimide reaction

Ling Xie, Wan Xia Li, Vincent A Barnett, Mark Schoenberg

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Previous assertions about the effect of alkylation of SIll and SH2 on the myosin high-salt calcium and EDTA ATPases have been summarized, and a simple procedure for obtaining the fractional labeling of SIll and SH2 after treatment of myosin with alkylating agents has been derived. A simple graphical procedure for illustrating the degree of preference of a particular alkylating agent for SIll over SH2 has also been developed. The procedures we developed were validated by applying them to two previously studied compounds, 4-(2-iodoacetamido)-TEMPO and 2,4-dinitrofluorobenzine, and then were used to determine a procedure for maximizing the extent of labeling of SIll alone by N-phenylmaleimide, a compound not previously studied in this manner. It was found that ~80% of the SIll sites could be alkylated without significant alkylation of SH2.

Original languageEnglish (US)
Pages (from-to)858-865
Number of pages8
JournalBiophysical journal
Volume72
Issue number2 I
DOIs
StatePublished - Feb 1997

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