Unique Behavior of Bacterially Expressed Rat Carnitine Palmitoyltransferase 2 and Its Catalytic Activity

DOI Web Site 参考文献26件
  • Akieda Kiri
    Institute of Advanced Medical Sciences, Tokushima University Graduate School of Pharmaceutical Sciences, Tokushima University
  • Takegawa Kazuto
    Institute of Advanced Medical Sciences, Tokushima University Graduate School of Pharmaceutical Sciences, Tokushima University
  • Ito Takeshi
    Institute of Advanced Medical Sciences, Tokushima University Graduate School of Pharmaceutical Sciences, Tokushima University
  • Nagayama Gaku
    Institute of Advanced Medical Sciences, Tokushima University Graduate School of Pharmaceutical Sciences, Tokushima University
  • Yamazaki Naoshi
    Graduate School of Pharmaceutical Sciences, Tokushima University
  • Nagasaki Yuka
    Awa Support Center, Tokushima University
  • Nishino Kohei
    Fujii Memorial Institute of Medical Sciences, Institute of Advanced Medical Sciences, Tokushima University
  • Kosako Hidetaka
    Fujii Memorial Institute of Medical Sciences, Institute of Advanced Medical Sciences, Tokushima University
  • Shinohara Yasuo
    Institute of Advanced Medical Sciences, Tokushima University Graduate School of Pharmaceutical Sciences, Tokushima University

抄録

<p>Mammalian type 2 carnitine parmitoyltransferase (EC 2.3.1.21), abbreviated as CPT2, is an enzyme involved in the translocation of fatty acid into the mitochondrial matrix space, and catalyzes the reaction acylcarnitine + CoA = acyl-CoA + carnitine. When rat CPT2 was expressed in Escherichia coli, its behavior was dependent on the presence or absence of i) its mitochondrial localization sequence and ii) a short amino acid sequence thought to anchor it to the mitochondrial inner membrane: CPT2 containing both sequences behaved as a hydrophobic protein, while recombinant CPT2 lacking both regions behaved as a water soluble protein; if only one region was present, the resultant proteins were observed in both fractions. Because relatively few protein species could be obtained from bacterial lysates as insoluble pellets under the experimental conditions used, selective enrichment of recombinant CPT2 protein containing both hydrophobic sequences was easily achieved. Furthermore, when CPT2 enriched in insoluble fraction was resuspended in an appropriate medium, it showed catalytic activity typical of CPT2: it was completely suppressed by the CPT2 inhibitor, ST1326, but not by the CPT1 inhibitor, malonyl-CoA. Therefore, we conclude that the bacterial expression system is an effective tool for characterization studies of mammalian CPT2.</p>

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