The monocarboxylate carrier from rat liver mitochondria. Purification and kinetic characterization in a reconstituted system.
The monocarboxylate (pyruvate) carrier was extracted from rat liver mitochondria with Triton X-100 in the presence of asolectin and partially purified by chromatography on HTP. The HTP eluate reconstituted in liposomes was shown to catalyze active pyruvatein/acetoacetateout and acetoacetatein/pyruvateout counter-exchange. Kinetic characterization of the reconstituted pyruvate carrier was achieved by an original spectrophotometric method consisting of determination of substrate release from proteoliposomes with a coupled enzymatic assay.
Capuano
F
F
Institute of Medical Biochemistry and Chemistry, University of Bari, Italy.
Di Paola
M
M
Azzi
A
A
Papa
S
S
D000090:Acetoacetates / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D003373:Coumaric Acids / Q000494:pharmacology; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008930:Mitochondria, Liver / Q000032:analysis*; D008970:Molecular Weight; D027501:Monocarboxylic Acid Transporters; D009243:NAD / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D051381:Rats; D013053:Spectrophotometry
D000090:Acetoacetates; D002352:Carrier Proteins; D003373:Coumaric Acids; D008081:Liposomes; D027501:Monocarboxylic Acid Transporters; D011773:Pyruvates; D009243:NAD; C007175:alpha-cyano-4-hydroxycinnamate; C016635:acetoacetic acid; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase