New Method to Prepare Single-Headed Heavy Meromyosin with High Purity and a High Yield
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 tr...
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| Main Authors: | , , , |
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| Format: | text |
| Language: | English |
| Published: |
Institutional Knowledge at Singapore Management University
1989
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| Subjects: | |
| Online Access: | https://ink.library.smu.edu.sg/lkcsb_research/1884 https://jb.oxfordjournals.org/cgi/content/abstract/106/4/606 |
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| Institution: | Singapore Management University |
| Language: | English |
| Summary: | 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. |
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