Purification of the active mitochondrial tricarboxylate carrier by hydroxylapatite chromatography

FEBS Letters

Stipani, I.¹  Palmieri, F.¹ 

[1] Institute of Biochemistry, Faculty of Pharmacy and CNR Unit for the Study of Mitochondria and Bioenergetics, University of Bari, Via Amendola 165/A, 70126 Bari, Italy

关键词: Tricarboxylate carrier;    Mitochondria;    Hydroxylapatite;    Reconstitution;    Membrane transport;    BTA;    benzenetricarboxylic acid;    DPG;    cardiolipin;    HTP;    hydroxylapatite;    NEM;    N-ethylmaleimide;    PEP;    phosphoenolpyruvate;    SDS;    sodium dodecylsulfate;    SMP;    submitochondrial particles;   

DOI  :  10.1016/0014-5793(83)81023-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The mitochondrial tricarboxylate carrier has been extracted from rat liver mitochondria or SMP with Triton X-100, in the presence of 1,2,3-BTA and DPG, and partially purified by chromatography on HTP. The purified fraction, which also contains the ADP/ATP carrier and the phosphate carrier, after incorporation into liposomes catalyzes a 1,2,3-BTA-sensitive [¹⁴C]citrate/citrate exchange. The tissue and substrate specificity, the inhibitor sensitivity and the kinetic properties of citrate transport in liposomes are similar to those described for the citrate transport in mitochondria. The maximal rate of citrate exchange in the reconstituted system is 338 μmol·min⁻¹·g protein⁻¹, at 30°C and pH 7.0.

【 授权许可】

Unknown