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

doi:10.1016/0003-2697(91)90223-G

Analytical Biochemistry

Volume 194, Issue 2, 1 May 1991, Pages 231-236

https://doi.org/10.1016/0003-2697(91)90223-G Get rights and content

Abstract

A detailed methodology is presented of two reconstitution protocols for the (Ca²⁺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.

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Analytical Biochemistry

Abstract

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Cited by (40)

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The sarcoplasmic-endoplasmic reticulum Ca<sup>2+</sup>-ATPase (SERCA) is the likely molecular target for the acute toxicity of the brominated flame retardant hexabromocyclododecane (HBCD)

Commonly used ryanodine receptor activator, 4-chloro-m-cresol (4CmC), is also an inhibitor of SERCA Ca<sup>2+</sup> pumps

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Tetrabromobisphenol A (TBBPA), induces cell death in TM4 Sertoli cells by modulating Ca<sup>2+</sup> transport proteins and causing dysregulation of Ca<sup>2+</sup> homeostasis

Caveolin-3-anchored microdomains at the rabbit sarcoplasmic reticulum membranes