New Method to Prepare Single-Headed Heavy Meromyosin with High Purity and a High Yield

Publication Type

Journal Article

Publication Date

1989

Abstract

We have developed a new method to prepare single-headed heavy meromyosin with high purity and a high yield. To examine whether the two heads on the same myosin molecule work cooperatively or not, it is important to prepare pure single-headed heavy mero-myosin. Myosin was extracted from myofibrils treated with a solution containing CyDTA, a strong divalent cation chelator. CyDTA treatment was essential to the production of sHMM. Then such myosin was digested with chymotrypsin in the presence of divalent cations at high ionic strength. Crude sHMM was separated from double-headed HMM by affinity chromatography using an ADP-column. Contaminating S1 was removed by gel filtration. Heavy chain of sHMM obtained by the present method had no nick. Purified sHMM showed normal EDTA-ATPase and Ca- ATPase. It interacted with thin filament and its ATPase was activated by actin normally.

Discipline

Physical Sciences and Mathematics

Research Areas

Quantitative Finance

Publication

Journal of Biochemistry

Volume

106

Issue

4

First Page

606

Last Page

661

ISSN

0021-924X

Identifier

10.1093/oxfordjournals.jbchem.a122903

Additional URL

https://doi.org/10.1093/oxfordjournals.jbchem.a122903

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