title: Nardilysin facilitates complex formation between mitochondrial malate dehydrogenase and citrate synthase.
abstract: Gel filtration chromatography showed that nardilysin activity in a rat testis or rat brain extract exhibited an apparent molecular weight of approximately 300 kDa compared to approximately 187 kDa for the purified enzyme. The addition of purified nardilysin to a rat brain extract, but not to an E. coli extract, produced the higher molecular species. The addition of a GST fusion protein containing the acidic domain of nardilysin eliminated the higher molecular weight nardilysin forms, suggesting that oligomerization involves the acidic domain of nardilysin. Using an immobilized nardilysin column, mitochondrial malate dehydrogenase (mMDH) and citrate synthase (CS) were isolated from a fractionated rat brain extract. Porcine mMDH, but not porcine cytosolic MDH, was shown to form a heterodimer with nardilysin. Mitochondrial MDH increased nardilysin activity about 50%, while nardilysin stabilized mMDH towards heat inactivation. CS was co-immunoprecipitated with mMDH only in the presence of nardilysin showing that nardilysin facilitates complex formation.
authors.affiliation: Department of Molecular and Cellular Biochemistry, University of Kentucky, Chandler Medical Center, 800 Rose Street, Lexington, KY 40536-0298, United States. mchow1@uky.edu
authors.forename: K Martin
authors.initials: KM
authors.lastname: Chow
authors.forename: Zhangliang
authors.initials: Z
authors.lastname: Ma
authors.forename: Jian
authors.initials: J
authors.lastname: Cai
authors.forename: William M
authors.initials: WM
authors.lastname: Pierce
authors.forename: Louis B
authors.initials: LB
authors.lastname: Hersh
mesh_terms: D000818:Animals; D002950:Citrate (si)-Synthase / Q000737:chemistry*; D008291:Malate Dehydrogenase / Q000737:chemistry*; D008297:Male; D008666:Metalloendopeptidases / Q000737:chemistry / Q000502:physiology*; D008928:Mitochondria / Q000201:enzymology*; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats
chemical_list: D008291:Malate Dehydrogenase; D002950:Citrate (si)-Synthase; D008666:Metalloendopeptidases; C087976:nardilysin