Methods of reconstitution of the purified sarcoplasmic reticulum (Ca2+Mg2+)-ATPase using bile salt detergents to form membranes of defined lipid to protein ratios or sealed vesicles
✍ Scribed by Francesco Michelangeli; Felix M. Munkonge
- Publisher
- Elsevier Science
- Year
- 1991
- Tongue
- English
- Weight
- 690 KB
- Volume
- 194
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A detailed methodology is presented of two reconstitution protocols for the (Ca(2+)-Mg2+)-ATPase from rabbit skeletal muscle, using the detergent potassium cholate. Method A was shown to produce fully fragmented membranes of definable lipid to protein ratios, which were unable to take up calcium upon hydrolysis of ATP. This protocol was shown to produce a homologous population of membranes with respect to their lipid and protein composition at lipid to protein ratios up to 900:1 (mol/mol). Method B produced vesicles only of high lipid to protein ratios (3000:1), which have the ability to accumulate calcium on addition of ATP. Calcium accumulation and ATP hydrolysis for the ATPase reconstituted into different fatty acyl chain length phospholipids were also studied.