New Method to Prepare Single-Headed Heavy Meromyosin with High Purity and a High Yield
著者
和文:
Kimiko Saeki,
徳永 万喜洋,
Hian Ann Ting,
Takeyuki Wakabayashi.
英文:
Kimiko Saeki,
Makio Tokunaga,
Hian Ann Ting,
Takeyuki Wakabayashi.
言語
English
掲載誌/書名
和文:
英文:
J. Biochem
巻, 号, ページ
Vol. 106
pp. 606-611
出版年月
1989年10月
出版者
和文:
英文:
会議名称
和文:
英文:
開催地
和文:
英文:
アブストラクト
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 meromyosin. 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.
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