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PMID: 2597117 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

The binding of Ca2+ ions to pig heart NAD+-isocitrate dehydrogenase and the 2-oxoglutarate dehydrogenase complex.

The Biochemical journal ·Vol. 263 ·No. 2 ·1989-10-15 ·Pages 453-62

Rutter GA, Denton RM

Abstract

1. The binding of Ca2+ ions to purified pig heart NAD+-isocitrate dehydrogenase and 2-oxoglutarate dehydrogenase, freed of contaminating Ca2+ by parvalbumin/polyacrylamide chromatography, has been studied by flow dialysis and by the use of fura-2. 2. For the 2-oxoglutarate dehydrogenase complex, 3.5 mol of Ca2+-binding sites/mol of complex were apparent, with an apparent dissociation constant (Kd value) for Ca2+ of 2.0 microM. These values were little affected by Mg2+ ions, ADP or 2-oxoglutarate. 3. By contrast, binding of Ca2+ to NAD+-isocitrate dehydrogenase (Kd = 14 microM) required ADP, isocitrate and Mg2+ ions. The number of Ca2+-binding sites associated with NAD+-isocitrate dehydrogenase was then 0.9 mol/mol of tetrameric enzyme. 4. The 2-oxoglutarate dehydrogenase complex bound ADP (as ADP3-) to a group of tight-binding sites (Kd = 3.1 microM) with a stoichiometry, 3.3 mol/mol of complex, similar to that for the binding of Ca2+; a variable number of much weaker sites (Kd = 100 microM) for ADP3- was also apparent.

MeSH Terms
Adenosine Diphosphate/metabolism,pharmacology Animals Benzofurans Binding Sites Calcium/metabolism Calmodulin/metabolism Chromatography Dialysis Fluorescent Dyes Fura-2 Isocitrate Dehydrogenase/metabolism Ketoglutarate Dehydrogenase Complex/metabolism Ketone Oxidoreductases/metabolism Magnesium/pharmacology Mitochondria, Heart/enzymology NAD Swine
Chemicals
Benzofurans Calmodulin Fluorescent Dyes NAD Adenosine Diphosphate Isocitrate Dehydrogenase Ketone Oxidoreductases Ketoglutarate Dehydrogenase Complex Magnesium Calcium Fura-2
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rutter G A
Department of Biochemistry, School of Medical Sciences, University of Bristol, U.K.
Denton R M
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2630453
Published
1989-10-15
Pages
453-62
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1133450
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2630453 PMC Full Text View journal details
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