Open AccessPartial Purification and Reconstitution of the α-Ketoglutarate Carrier from Corn (Zea mays L.) Mitochondria
Author(s) -
Giuseppe Genchi,
Adelmo De Santis,
Carmela Ponzone,
Ferdinando Palmieri
Publication year - 1991
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.96.4.1003
Subject(s) - cardiolipin , zea mays , chromatography , biochemistry , chemistry , mitochondrion , specific activity , elution , biology , enzyme , membrane , phospholipid , agronomy
The alpha-ketoglutarate carrier from corn shoot mitochondria (Zea mays L., B 73) was solubilized in Triton X-114 and partially purified by chromatography on hydroxyapatite and celite in the presence of cardiolipin. On SDS-gel electrophoresis, the hydroxyapatite/celite eluate showed various protein bands between 12 and 70 kilodaltons. When reconstituted into liposomes, the alpha-ketoglutarate transport protein catalyzed a phthalonate-sensitive alpha-ketoglutarate/alpha-ketoglutarate exchange. The protein was purified 60-fold with a recovery of 88% with respect to the mitochondrial extract. The protein yield was 0.6%. The properties of the reconstituted carrier, i.e. requirement for a counter-anion, substrate specificity, and inhibitor sensitivity, were similar to those of the alpha-ketoglutarate transport system as characterized in plant and animal mitochondria.