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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
1990-10-3
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pubmed:abstractText |
Bovine cytochrome c oxidase subunits were separated by reverse phase high performance liquid chromatography using a C4 column eluted with water and an acetonitrile gradient, both containing 0.1% trifluoroacetic acid. Subunits I and III precipitated in this solvent and could not be analyzed; the remaining eleven subunits were dissociated, denatured, soluble and could be resolved by elution from the column. The protein subunit eluting in each chromatographic peak was identified by a combination of polyacrylamide gel electrophoresis in sodium dodecyl sulfate, NH2-terminal amino acid sequencing, and amino acid analysis. Each subunit produced a single elution peak with the exception of subunit VIc (nomenclature of Kadenbach et al., 1983, Anal. Biochem. 129, 517-521), which eluted from the column as two well-resolved peaks. Sequence analysis showed that the two subunit VIc elution peaks resulted from partial chemical blockage of the alpha-amino serine residue of subunit VIc. The C4 reverse phase HPLC was used to document specific subunit removal from bovine cytochrome c oxidase either by tryptic digestion or by dodecyl maltoside extraction. The described HPLC method for separating cytochrome c oxidase subunits should be applicable for the analysis of other multisubunit proteins, especially other multisubunit membrane protein complexes.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0003-9861
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
281
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
239-44
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:2168156-Amino Acid Sequence,
pubmed-meshheading:2168156-Amino Acids,
pubmed-meshheading:2168156-Animals,
pubmed-meshheading:2168156-Cattle,
pubmed-meshheading:2168156-Chromatography, High Pressure Liquid,
pubmed-meshheading:2168156-Electron Transport Complex IV,
pubmed-meshheading:2168156-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:2168156-Macromolecular Substances,
pubmed-meshheading:2168156-Molecular Sequence Data
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pubmed:year |
1990
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pubmed:articleTitle |
Subunit analysis of bovine cytochrome c oxidase by reverse phase high performance liquid chromatography.
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pubmed:affiliation |
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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