Kinetics of the Reconstituted Tricarboxylate Carrier from Eel Liver Mitochondria View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1998-12

AUTHORS

V. Zara, L. Palmieri, M.R. Franco, M. Perrone, G.V. Gnoni, F. Palmieri

ABSTRACT

The tricarboxylate carrier from eel liver mitochondria was purified by chromatography on hydroxyapatite and Matrix Gel Blue B and reconstituted into liposomes by removal of the detergent with Amberlite. Optimal transport activity was obtained by using a phospholipid concentration of 11.5 mg/ml, a Triton X- 114/phospholipid ratio of 0.9, and ten passages through the same Amberlite column. The activity of the carrier was influenced by the phospholipid composition of the liposomes, being increased by cardiolipin and phosphatidylethanolamine and decreased by phosphatidylinositol. The reconstituted tricarboxylate carrier catalyzed a first-order reaction of citrate/citrate or citrate/malate exchange. The maximum transport rate of external [14C]citrate was 9.0 mmol/min per g of tricarboxylate carrier protein at 25 degrees C and this value was virtually independent of the type of substrate present in the external or internal space of the liposomes. The half-saturation constant (Km) was 62 microM for citrate and 541 microM for malate. The activation energy of the citrate/citrate exchange reaction was 74 kJ/mol from 5 to 19 degrees C and 31 kJ/mol from 19 to 35 degrees C. The rate of the exchange had an external pH optimum of 8. More... »

PAGES

555-563

Identifiers

URI

http://scigraph.springernature.com/pub.10.1023/a:1020532500749

DOI

http://dx.doi.org/10.1023/a:1020532500749

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1022014038

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/10206475


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