Molecular identification and functional characterization of a novel glutamate transporter in yeast and plant mitochondria

Vito Porcelli, Angelo Vozza, Valeria Calcagnile, Ruggiero Gorgoglione, Roberto Arrigoni, Flavia Fontanesi, Carlo M.T. Marobbio, Alessandra Castegna, Ferdinando Palmieri, Luigi Palmieri

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The genome of Saccharomyces cerevisiae encodes 35 members of the mitochondrial carrier family (MCF) and 58 MCF members are coded by the genome of Arabidopsis thaliana, most of which have been functionally characterized. Here two members of this family, Ymc2p from S. cerevisiae and BOU from Arabidopsis, have been thoroughly characterized. These proteins were overproduced in bacteria and reconstituted into liposomes. Their transport properties and kinetic parameters demonstrate that Ymc2p and BOU transport glutamate, and to a much lesser extent L-homocysteinesulfinate, but not other amino acids and many other tested metabolites. Transport catalyzed by both carriers was saturable, inhibited by mercuric chloride and dependent on the proton gradient across the proteoliposomal membrane. The growth phenotype of S. cerevisiae cells lacking the genes ymc2 and agc1, which encodes the only other S. cerevisiae carrier capable to transport glutamate besides aspartate, was fully complemented by expressing Ymc2p, Agc1p or BOU. Mitochondrial extracts derived from ymc2Δagc1Δ cells, reconstituted into liposomes, exhibited no glutamate transport at variance with wild-type, ymc2Δ and agc1Δ cells, showing that S. cerevisiae cells grown in the presence of acetate do not contain additional mitochondrial transporters for glutamate besides Ymc2p and Agc1p. Furthermore, mitochondria isolated from wild-type, ymc2Δ and agc1Δ strains, but not from the double mutant ymc2Δagc1Δ strain, swell in isosmotic ammonium glutamate showing that glutamate is transported by Ymc2p and Agc1p together with a H+. It is proposed that the function of Ymc2p and BOU is to transport glutamate across the mitochondrial inner membrane and thereby play a role in intermediary metabolism, C1 metabolism and mitochondrial protein synthesis.

Original languageEnglish (US)
JournalBiochimica et Biophysica Acta - Bioenergetics
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Amino Acid Transport System X-AG
Mitochondria
Yeast
Glutamic Acid
Yeasts
Saccharomyces cerevisiae
Genes
Arabidopsis
Metabolism
Liposomes
Genome
Membranes
Mercuric Chloride
Mitochondrial Proteins
Mitochondrial Membranes
Metabolites
Ammonium Compounds
Kinetic parameters
Aspartic Acid
Transport properties

Keywords

  • A bout de souffle
  • C1 metabolism
  • Mitochondrial carriers
  • Mitochondrial glutamate carrier
  • Photorespiration
  • YMC2

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Molecular identification and functional characterization of a novel glutamate transporter in yeast and plant mitochondria. / Porcelli, Vito; Vozza, Angelo; Calcagnile, Valeria; Gorgoglione, Ruggiero; Arrigoni, Roberto; Fontanesi, Flavia; Marobbio, Carlo M.T.; Castegna, Alessandra; Palmieri, Ferdinando; Palmieri, Luigi.

In: Biochimica et Biophysica Acta - Bioenergetics, 01.01.2018.

Research output: Contribution to journalArticle

Porcelli, Vito ; Vozza, Angelo ; Calcagnile, Valeria ; Gorgoglione, Ruggiero ; Arrigoni, Roberto ; Fontanesi, Flavia ; Marobbio, Carlo M.T. ; Castegna, Alessandra ; Palmieri, Ferdinando ; Palmieri, Luigi. / Molecular identification and functional characterization of a novel glutamate transporter in yeast and plant mitochondria. In: Biochimica et Biophysica Acta - Bioenergetics. 2018.
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AU - Vozza, Angelo

AU - Calcagnile, Valeria

AU - Gorgoglione, Ruggiero

AU - Arrigoni, Roberto

AU - Fontanesi, Flavia

AU - Marobbio, Carlo M.T.

AU - Castegna, Alessandra

AU - Palmieri, Ferdinando

AU - Palmieri, Luigi

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KW - A bout de souffle

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KW - Mitochondrial glutamate carrier

KW - Photorespiration

KW - YMC2

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