The Tricarboxylate Carrier

  • A. Azzi
  • M. Glerum
  • R. Koller
  • W. Mertens
Conference paper
Part of the NATO ASI Series book series (volume 83)

Summary

The citrate carrier has recently been purified from rat liver mitochondria by several groups using different methods. A 37–38 kDa protein has been prepared by Silica Gel 60 chromatography by our group (Claeys and Azzi, 1989; Glerum et al., 1990). The specific citrate transport activity of this preparation is not significantly different from that measured in mitochondria and it is inhibitable by 1, 2, 3- benzenetricarboxylic acid. Bisaccia et al. (1990) have reported the isolation of a 30 kDa protein by Celite 535 chromatography, and Kaplan et al. (1990) have isolated a 32.5 kDa protein by Matrex Orange, Matrex Blue and Affi-Gel chromatography. The clarification of the structural diversities of the above-described preparations has been one of the objectives of the present work. Peptide mapping, after cleavage of the different protein preparations with Endoproteinase Glu-C (Staphylococcus aureus V8 protease), cyanogen bromide and hydroxylamine, has failed to support any structural homologies. The 38 kD protein is N-terminally blocked. The peptides obtained from enodproteinase Glu-C or hydroxylamine lysis, after separation on SDS-PAGE, and those obtained after CNBr cleavage and HPLC purification have been partially sequenced. The protein sequence information is being used to obtain a full-length cDNA, which will allow further characterisation of the carrier at the molecular level.

Keywords

Cholesterol Citrate Electrophoresis Fractionation Polypeptide 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1994

Authors and Affiliations

  • A. Azzi
    • 1
  • M. Glerum
    • 1
  • R. Koller
    • 1
  • W. Mertens
    • 1
  1. 1.Institut für Biochemie und MolekularbiologieUniversität BernBernSwitzerland

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