Nardilysin facilitates complex formation between mitochondrial malate dehydrogenase and citrate synthase

K. Martin Chow; Zhangliang Ma; Jian Cai; William M. Pierce; Louis B. Hersh

doi:10.1016/j.bbagen.2005.02.010

Nardilysin facilitates complex formation between mitochondrial malate dehydrogenase and citrate synthase

K. Martin Chow, Zhangliang Ma, Jian Cai, William M. Pierce, Louis B. Hersh

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

Gel filtration chromatography showed that nardilysin activity in a rat testis or rat brain extract exhibited an apparent molecular weight of ∼300 kDa compared to ∼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.

Original language	English
Pages (from-to)	292-301
Number of pages	10
Journal	Biochimica et Biophysica Acta - General Subjects
Volume	1723
Issue number	1-3
DOIs	https://doi.org/10.1016/j.bbagen.2005.02.010
State	Published - May 25 2005

Bibliographical note

Funding Information:
This research was supported by NIH/NIDA grant DA 02243. The University of Louisville Biomolecular Mass Spectrometry Laboratory is supported in part by NIH Grant 1S10RR11368-01A1 (W.M.P.), the State of Kentucky Physical Facilities Trust Fund, the University of Louisville, School of Medicine, and the University of Louisville Research Foundation. We would also like to thank Dr. E. DeMoll (Department of Chemistry, University of Kentucky) for assisting in the IEF analysis.

Keywords

Acidic domain
Centaurin-alpha
Metabolon
N-arginine dibasic convertase
Protein-protein interaction
Tyrosyl-tRNA ligase

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/j.bbagen.2005.02.010

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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 ∼300 kDa compared to ∼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.",

keywords = "Acidic domain, Centaurin-alpha, Metabolon, N-arginine dibasic convertase, Protein-protein interaction, Tyrosyl-tRNA ligase",

author = "Chow, {K. Martin} and Zhangliang Ma and Jian Cai and Pierce, {William M.} and Hersh, {Louis B.}",

note = "Funding Information: This research was supported by NIH/NIDA grant DA 02243. The University of Louisville Biomolecular Mass Spectrometry Laboratory is supported in part by NIH Grant 1S10RR11368-01A1 (W.M.P.), the State of Kentucky Physical Facilities Trust Fund, the University of Louisville, School of Medicine, and the University of Louisville Research Foundation. We would also like to thank Dr. E. DeMoll (Department of Chemistry, University of Kentucky) for assisting in the IEF analysis.",

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AU - Chow, K. Martin

AU - Ma, Zhangliang

AU - Cai, Jian

AU - Pierce, William M.

AU - Hersh, Louis B.

N1 - Funding Information: This research was supported by NIH/NIDA grant DA 02243. The University of Louisville Biomolecular Mass Spectrometry Laboratory is supported in part by NIH Grant 1S10RR11368-01A1 (W.M.P.), the State of Kentucky Physical Facilities Trust Fund, the University of Louisville, School of Medicine, and the University of Louisville Research Foundation. We would also like to thank Dr. E. DeMoll (Department of Chemistry, University of Kentucky) for assisting in the IEF analysis.

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N2 - Gel filtration chromatography showed that nardilysin activity in a rat testis or rat brain extract exhibited an apparent molecular weight of ∼300 kDa compared to ∼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.

AB - Gel filtration chromatography showed that nardilysin activity in a rat testis or rat brain extract exhibited an apparent molecular weight of ∼300 kDa compared to ∼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.

KW - Acidic domain

KW - Centaurin-alpha

KW - Metabolon

KW - N-arginine dibasic convertase

KW - Protein-protein interaction

KW - Tyrosyl-tRNA ligase

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Nardilysin facilitates complex formation between mitochondrial malate dehydrogenase and citrate synthase

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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