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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1998-10-2
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pubmed:abstractText |
The carnitine/acylcarnitine carrier from rat liver mitochondria was overexpressed in Escherichia coli. The expressed protein, recovered as inclusion bodies, was solubilized with sarkosyl and purified by Sephadex G-200 and celite chromatography. A yield of 15 mg of purified transport protein per liter of cell culture was obtained. Upon reconstitution into liposomes, the purified carrier catalyzed a [3H]carnitine/carnitine exchange inhibited by maleimides, mercurials, and sulfobetaines. Carnitine esters of various lengths were also transported. The Km for carnitine uptake was 0.47 +/- 0.11 mM, the Vmax of the exchange was 0.78 +/- 0.24 mmol/min per gram of protein, and the Ki for octanoylcarnitine was 13.5 +/- 4.3 microM. The transport properties of the recombinant carrier were virtually identical to those of the native transporter. These studies represent the first overexpression of the functionally active mitochondrial carnitine/acylcarnitine carrier, thus enabling structure/function analysis of this protein by site-directed mutagenesis.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carnitine,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Maleimides,
http://linkedlifedata.com/resource/pubmed/chemical/Organomercury Compounds,
http://linkedlifedata.com/resource/pubmed/chemical/Proteolipids,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/proteoliposomes
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0006-291X
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
28
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pubmed:volume |
249
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
589-94
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:9731180-Animals,
pubmed-meshheading:9731180-Binding Sites,
pubmed-meshheading:9731180-Biological Transport, Active,
pubmed-meshheading:9731180-Carnitine,
pubmed-meshheading:9731180-Carrier Proteins,
pubmed-meshheading:9731180-Escherichia coli,
pubmed-meshheading:9731180-Gene Expression,
pubmed-meshheading:9731180-Kinetics,
pubmed-meshheading:9731180-Maleimides,
pubmed-meshheading:9731180-Mitochondria, Liver,
pubmed-meshheading:9731180-Organomercury Compounds,
pubmed-meshheading:9731180-Proteolipids,
pubmed-meshheading:9731180-Rats,
pubmed-meshheading:9731180-Recombinant Proteins
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pubmed:year |
1998
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pubmed:articleTitle |
Bacterial overexpression, purification, and reconstitution of the carnitine/acylcarnitine carrier from rat liver mitochondria.
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pubmed:affiliation |
Department of Pharmaco-Biology, University of Bari, Italy.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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