title: Reaction mechanism of the reconstituted tricarboxylate carrier from rat liver mitochondria.
abstract: Transport of citrate and malate by the tricarboxylate carrier from rat liver mitochondria has been studied in a reconstituted system. Homologous citrate/citrate antiport and heterologous (electroneutral) citrate/malate antiport was kinetically analyzed. The maximal rates of the two exchange modes did not vary significantly within pH 7.0 to 7.8 which is the optimum pH-range for transport activity. On the other hand, the apparent transport affinity varied considerably within this range. Calculations on the basis of the different pK values for citrate and malate indicate that only H-citrate2- and malate2- are accepted as transport species by the tricarboxylate carrier. A complete set of half-saturation constants was established for citrate and malate on both the external and the internal side of the membrane. Both the Km and Vmax for citrate and malate were independent of the nature of the countersubstrate at the other side of the membrane. Bisubstrate initial velocity analyses of the exchange reaction resulted in a kinetic pattern which is consistent with a sequential antiport mechanism. This type of mechanism implies formation of a ternary complex of the carrier with two substrate molecules before the transport reaction occurs. Thus the tricarboxylate carrier falls into the functional family of mitochondrial carrier proteins showing sequential transport mechanisms.
authors.affiliation: Department of Pharmaco-Biology, University of Bari, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Bisaccia
authors.forename: A
authors.initials: A
authors.lastname: De Palma
authors.forename: T
authors.initials: T
authors.lastname: Dierks
authors.forename: R
authors.initials: R
authors.lastname: Krämer
authors.forename: F
authors.initials: F
authors.lastname: Palmieri
mesh_terms: D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008930:Mitochondria, Liver / Q000737:chemistry* / Q000378:metabolism; D051381:Rats
chemical_list: D002352:Carrier Proteins; C020723:citrate-binding transport protein