Effect of aspartate and glutamate on the oxoglutarate carrier investigated in rat heart mitochondria and inverted submitochondrial vesicles.
Interaction of glutamate and aspartate with the oxoglutarate carrier was investigated in rat heart mitochondria or inverted submitochondrial particles. With mitochondria, glutamate and aspartate had no effect on the initial rate of oxoglutarate or malate uptake. With inverted submitochondrial vesicles, binding experiments indicated that aspartate bound to the oxoglutarate carrier on its matricial face and increased the affinity of the substrate binding site for malate but did not change the affinity for oxoglutarate. Glutamate had no effect on both substrate bindings. The dissociation constants of the binary substrate-carrier complexes on the matricial side were determined (1.28 +/- 0.15 mM for oxoglutarate and 2.22 +/- 0.26 mM for malate). These values, compared with those obtained previously on the cytosolic side of intact mitochondria, confirmed the asymmetry of the carrier in the native membrane (higher affinities on the cytosolic face). It is concluded that (1) aspartate and glutamate are not cytosolic effectors of the oxoglutarate carrier, (2) matricial aspartate is a positive effector of the binding of malate on the matricial side of the oxoglutarate carrier, and (3) such a characteristic may play a role in the regulation of the oxoglutarate carrier. Thus, it may be emphasized that (1) this observation is the first clear evidence of a well-defined 'sophisticated regulation' (allosteric) of a mitochondrial metabolite carrier, and (2) this regulation of the oxoglutarate carrier may have important consequences on the efficiency of reducing equivalent import in the matrix space by the malate-aspartate shuttle.
Hautecler
J J
JJ
Laboratoire de Bioénergétique, Université de Liège, Belgium.
Sluse-Goffart
C M
CM
Evens
A
A
Duyckaerts
C
C
Sluse
F E
FE
D000818:Animals; D001224:Aspartic Acid / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism*; D005971:Glutamates / Q000494:pharmacology*; D018698:Glutamic Acid; D007656:Ketoglutaric Acids / Q000378:metabolism; D007700:Kinetics; D008293:Malates / Q000378:metabolism; D026901:Membrane Transport Proteins; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000378:metabolism; D051381:Rats
D002352:Carrier Proteins; D005971:Glutamates; D007656:Ketoglutaric Acids; D008293:Malates; D026901:Membrane Transport Proteins; C021723:oxoglutarate translocator; D001224:Aspartic Acid; D018698:Glutamic Acid; C030298:malic acid