Resolution and reconstitution of active transport of calcium by a protein(s) from Mycobacterium phlei

Membrane protein(s) responsible for the active transport of calcium in membrane vesicles from Mycobacterium phlei have been solubilized from membranes by sodium cholate treatment and partially purified using a hydrophobic resin. Reconstitution of calcium transport was demonstrated by reconstitution...

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Bibliographic Details
Published in:The Journal of biological chemistry 1979-08, Vol.254 (15), p.6861-6864
Main Authors: Lee, S H, Kalra, V K, Brodie, A F
Format: Article
Language:English
Subjects:
Biological Transport, Active - drug effects
Calcimycin - pharmacology
Calcium - metabolism
Carbonyl Cyanide m-Chlorophenyl Hydrazone - pharmacology
Cell Membrane - metabolism
Kinetics
Lanthanum - pharmacology
Liposomes
Membrane Proteins - isolation & purification
Membrane Proteins - metabolism
Mycobacterium - metabolism
Mycobacterium phlei - metabolism
Proteolipids
Terbium - pharmacology
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Summary:Membrane protein(s) responsible for the active transport of calcium in membrane vesicles from Mycobacterium phlei have been solubilized from membranes by sodium cholate treatment and partially purified using a hydrophobic resin. Reconstitution of calcium transport was demonstrated by reconstitution of detergent extracted membranes with the partially purified protein. The uptake of calcium in the reconstituted system was sensitive to proton-conducting uncouplers. Liposomes prepared with partially purified calcium translocating protein were capable of accumulating calcium. The uptake of calcium in this system occurred as a result of an artificial proton gradient generated by the reduction of entrapped ferricyanide with ascorbate-benzoquinone serving as a hydrogen carrier. The addition of the ionophore A23187 caused efflux of accumulated calcium in both native and proteoliposomal-reconstituted system.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)50252-0