Functional reconstitution of the partially purified aspartate-glutamate carrier from mitochondria |
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| Authors: | A.D.M. van Mansfeld H.A.A.M. van Teeffelen P.D. Baas G.H. Veeneman J.H. van Boom H.S. Jansz |
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| Affiliation: | 1. Institute of Molecular Biology and Laboratory for Physiological Chemistry, State University of Utrecht, Padualaan 8, 3584 CH UtrechtThe Netherlands;2. Department of Organic Chemistry, State University of Leiden, 2300 RA Leiden, The Netherlands |
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| Abstract: | The aspartate/glutamate carrier from beef heart mitochondria has been solubilized with detergent. The transport protein was partially purified by chromatography on hydroxyapatite in the presence of dodecyl octaoxyethylene ether and high concentrations of ammonium acetate. During purification, the aspartate/glutamate carrier was identified by functional reconstitution into egg yolk phospholipid liposomes. After hydroxyapatite chromatography the protein is 30 fold enriched in aspartate/glutamate transport activity but still contains ADP/ATP-carrier and phosphate carrier. The reconstituted activity is specific for exchange of L-aspartate and L-glutamate and is similar to intact mitochondria with respect to substrate affinity and inhibitor sensitivity. |
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| Keywords: | Aspartate/glutamate carrier Beef heart mitochondria Liposome Reconstitution Detergent Kinetics asp aspartate BKA bongkrekate CAT carboxyatractylate glu glutamate PalP pyridoxalphosphate Pipes |
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