Isolation and functional reconstitution of the aspartate/glutamate carrier from mitochondria

Krämer, Reinhard; Kürzinger, Gaby; Heberger, Claudia

doi:10.1016/0003-9861(86)90063-9

Cited by 29 publications

(6 citation statements)

References 19 publications

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“…Since the purity of the reconstituted protein fraction is still unknown, the specific activity of carnitine exchange obtained in proteoliposomes cannot be correlated quantitatively to the original mitochondrial activity. However, the rate of reconstituted carnitine/carnitine exchange is in the same order of magnitude as the values found for other reconstituted mitochondrial metabolite carriers [18,19] and definitely lower than the rate of the reconstituted phosphate carrier [20] which also has a higher specific activity in mitochondria.…”

Section: Properties Of the Reconstituted Carnitine Carriersupporting

confidence: 56%

Purification of the mitochondrial carnitine carrier by chromatography on hydroxyapatite and celite

Indiveri

Palmieri

1989

FEBS Letters

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The carnitine carrier from rat liver mitochondria has been extracted with Triton X-100 and partially purified by chromatography on hydroxyapatite and celite. During purification the activity of the carrier was monitored by functional reconstitution into liposomes. The purified fraction is 250-fold enriched with respect to the N-ethylmaleimide-sensitive carnitine/carnitine transport activity. The substrate specificity and the inhibitor sensitivity of carnitine transport in liposomes resemble closely those described for the transport of carnitine in mitochondria.

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Section: Properties Of the Reconstituted Carnitine Carriersupporting

confidence: 56%

Purification of the mitochondrial carnitine carrier by chromatography on hydroxyapatite and celite

Indiveri

Palmieri

1989

FEBS Letters

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“…Knowledge about the mitochondrial AGC has existed for a long time, and its main properties have been studied in intact mitochondria (for references see LaNoue and Schoolwerth, 1979) and in reconstituted proteoliposomes (Krämer et al ., 1986; Dierks and Krämer, 1988; Dierks et al ., 1988; Bisaccia et al ., 1992). However, the protein responsible for the exchange had not been identified hitherto.…”

Section: Discussionmentioning

confidence: 99%

Citrin and aralar1 are Ca2+-stimulated aspartate/glutamate transporters in mitochondria

Palmieri

Pardo

Lasorsa

et al. 2001

EMBO J

458

410

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The mitochondrial aspartate/glutamate carrier catalyzes an important step in both the urea cycle and the aspartate/malate NADH shuttle. Citrin and aralar1 are homologous proteins belonging to the mitochondrial carrier family with EF-hand Ca 2+ -binding motifs in their N-terminal domains. Both proteins and their C-terminal domains were overexpressed in Escherichia coli, reconstituted into liposomes and shown to catalyze the electrogenic exchange of aspartate for glutamate and a H + . Overexpression of the carriers in transfected human cells increased the activity of the malate/aspartate NADH shuttle. These results demonstrate that citrin and aralar1 are isoforms of the hitherto unidenti®ed aspartate/glutamate carrier and explain why mutations in citrin cause type II citrullinemia in humans. The activity of citrin and aralar1 as aspartate/glutamate exchangers was stimulated by Ca 2+ on the external side of the inner mitochondrial membrane, where the Ca 2+ -binding domains of these proteins are localized. These results show that the aspartate/glutamate carrier is regulated by Ca 2+ through a mechanism independent of Ca 2+ entry into mitochondria, and suggest a novel mechanism of Ca 2+ regulation of the aspartate/malate shuttle.

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“…The transport properties and functional roles of the AGCs aralar1 and citrin have been analysed [22,37,39,40,[84][85][86][87][88][89]. Their large molecular mass, about 70 kDa, matches that of partially purified AGC preparations from bovine heart mitochondria [37,39,82]. The transport properties of both carriers reconstituted into proteoliposomes fully match those of the native AGC regarding substrate specificity, transport affinities, inhibitor sensitivity and voltage dependence [22].…”

Section: Mcs For Glutamatementioning

confidence: 83%

New mitochondrial carriers: an overview

Arco

Satrústegui

2005

Cell. Mol. Life Sci.

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The transport of metabolites, nucleotides and cofactors across the mitochondrial inner membrane is performed by members of mitochondrial carrier family (MCF). These proteins share marked structural features that have made feasible the functional characterization of numerous MCs in the last years. The MCs responsible for transport activities in mitochondria known for decades such as glutamate uptake or ATP-Mg/Pi exchange have recently been identified as well as novel carriers such as those involved in S-adenosylmethionine or thiamine pyrophosphate uptake. Here, after a brief review of the novel data on structural characteristics and import mechanisms of MCF members, we present an exhaustive compilation of human MC sequences, including previously characterized carriers, together with their respective Saccharomyces cerevisiae orthologues, ordered according to the phylogenetic analysis of el Moualij and co-workers [Yeast (1997) 13: 573-581]. We have detected the existence of at least 49 human MC sequences, including those of yet unknown function. An overview of novel MCF members functionally characterized in recent years in mammals and in yeast genomes is presented.

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Isolation and functional reconstitution of the aspartate/glutamate carrier from mitochondria

Cited by 29 publications

References 19 publications

Purification of the mitochondrial carnitine carrier by chromatography on hydroxyapatite and celite

Purification of the mitochondrial carnitine carrier by chromatography on hydroxyapatite and celite

Citrin and aralar1 are Ca2+-stimulated aspartate/glutamate transporters in mitochondria

New mitochondrial carriers: an overview

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