The ? and ? subunits play distinct functional roles in the ?₂?? heterotetramer of human NAD-dependent isocitrate dehydrogenase.

ABSTRACT: Human NAD-dependent isocitrate dehydrogenase existing as the ?₂?? heterotetramer, catalyzes the decarboxylation of isocitrate into ?-ketoglutarate in the Krebs cycle, and is allosterically regulated by citrate, ADP and ATP. To explore the functional roles of the regulatory ? and ? subunits, we systematically characterized the enzymatic properties of the holoenzyme and the composing ?? and ?? heterodimers in the absence and presence of regulators. The biochemical and mutagenesis data show that ?? and ?? alone have considerable basal activity but the full activity of ?₂?? requires the assembly and cooperative function of both heterodimers. ?₂?? and ?? can be activated by citrate or/and ADP, whereas ?? cannot. The binding of citrate or/and ADP decreases the S_{0.5,isocitrate} and thus enhances the catalytic efficiencies of the enzymes, and the two activators can act independently or synergistically. Moreover, ATP can activate ?₂?? and ?? at low concentration and inhibit the enzymes at high concentration, but has only inhibitory effect on ??. Furthermore, the allosteric activation of ?₂?? is through the ? subunit not the ? subunit. These results demonstrate that the ? subunit plays regulatory role to activate the holoenzyme, and the ? subunit the structural role to facilitate the assembly of the holoenzyme.

SUBMITTER: Ma T 

PROVIDER: S-EPMC5282582 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature