A rapid assay for actin-associated high-affinity cytochalasin binding sites based on isoelectric precipitation of soluble protein
✍ Scribed by Diane Chang Lin; Shin Lin
- Publisher
- Elsevier Science
- Year
- 1980
- Tongue
- English
- Weight
- 574 KB
- Volume
- 103
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
When protein from a red cell ghost extract precipitates in the presence of ["Hldihydrocytochalasin B (a drug which inhibits cell motility in many eucaryotic cells), it carries along with it a saturable amount of the labeled drug. This phenomenon, which probably reflects the isoelectric precipitation of the cytochalasin:receptor complex, occurs at around pH 5, is independent of the type of acid used to adjust the pH, and is inhibited by high salt concentrations and by pretreatment of extract protein with chymotrypsin. A simple, rapid, and quantitative assay based on the isoelectric precipitation reaction has been devised. This assay was used to demonstrate the presence of actin-associated high-affinity binding sites for dihydrocytochalasin B in extracts of a variety of eucaryotic cells.