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2007 | 54 | 3 | 575-581
Article title

A new method to precipitate myosin v from rat brain soluble fraction

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EN
Abstracts
EN
Myosin can be precipitated from soluble fraction under different assay conditions. This paper describes a new method for precipitating myosin V from rat brain soluble fraction. Brains were homogenized in 50 mM imidazole/HCl buffer, pH 8.0, containing 10 mM EDTA/EGTA, 250 mM sucrose, 1 mM DTT and 1 mM benzamidine, centrifuged at 45000 × g for 40 min and the supernatant was frozen at -20 °C. Forty-eight hours later, the supernatant was thawed, centrifuged at 45000 × g for 40 min and the precipitate was washed in 20 mM imidazole buffer pH 8.0. SDS/PAGE analysis showed four polypeptides in the precipitate: 205, 150, 57 and 43 kDa. The precipitate presented high Mg2+-ATPase activity, which co-purifies with p205. This polypeptide was recognized by a specific myosin V antibody and was proteolised by calpain, generating two stable polypeptides: p130 and p90. The Mg2+-ATPase activity was not stimulated by calcium in both the absence and presence of exogenous calmodulin and the K+/EDTA-ATPase activity represented 25% of the Mg2+-ATPase activity. In this work, myosin V from rat brain was precipitated by freezing the soluble fraction and was co-purificated with a 45 kDa polypeptide.
Keywords
EN
Publisher

Year
Volume
54
Issue
3
Pages
575-581
Physical description
Dates
published
2007
received
2007-06-06
revised
2007-09-03
accepted
2007-09-12
(unknown)
2007-09-17
Contributors
  • Universidade Federal de Uberlandia, Institute of Genetics and Biochemistry, Uberlândia - MG, Brazil
  • Universidade Federal de Uberlandia, Institute of Genetics and Biochemistry, Uberlândia - MG, Brazil
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Document Type
Publication order reference
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YADDA identifier
bwmeta1.element.bwnjournal-article-abpv54p575kz
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