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Characterization of the alpha -ketoglutarate dehydrogenase complex from Fasciola hepatica: potential implications for the role of calcium in the regulation of helminth mitochondrial metabolism



Characterization of the alpha -ketoglutarate dehydrogenase complex from Fasciola hepatica: potential implications for the role of calcium in the regulation of helminth mitochondrial metabolism



Molecular and Biochemical Parasitology 81(2): 243-246



The sensitivity of the alpha -ketoglutarate dehydrogenase complex (KGDC) of adult Fasciola hepatica to calcium was investigated. The specific activities of purified KGDC ranged from 2.3 to 5.6 micro mol/minute/mg protein. Analysis of the KGDC by SDS-PAGE indicated the presence of 3 major bands of 107, 56 and 48 kDa with MWs similar to alpha -ketoglutarate dehydrogenase, dihydrolipoyl dehydrogenase and dihydrolipoyl transsuccinylase subunits of the mammalian complex. Furthermore, 2 breakdown products with minor bands of about 20 kDa were also detected. KGDC activity (based on apparent Km for alpha -ketoglutarate ( alpha -KG)) was not affected by the addition of calcium, EGTA, adenine nucleotides or amino acids such as proline, histidine, arginine, alanine, glutamine, and aspartic and glutamic acids. When the pH of the potassium phosphate assay buffer was decreased from 7.5 to 6.5, the apparent Km of alpha -KG decreased from 48.1 +or- 5.1 to 19.8 +or- 6.5 (n=3).

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Accession: 002775999

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PMID: 8898339


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