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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
1994-3-18
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pubmed:databankReference |
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pubmed:abstractText |
A cDNA clone encoding a hydrophilic protein with a calculated molecular mass of 13,839 Da was isolated by shotgun screening with an anti-V-ATPase holoenzyme serum. The deduced amino acid sequence showed no significant homology to any other known protein. Southern blots revealed the existence of only one gene encoding the 14-kDa protein. Monospecific antibodies purified by affinity to the recombinant protein demonstrated the presence of a 14-kDa protein in the highly purified goblet cell apical membrane and inhibited ATP-dependent proton transport as well as V-ATPase activity to the same extent. Thus, the 14-kDa protein was shown to be a part of the V-ATPase holoenzyme. Binding of the monospecific antibodies to the ATPase seemed to require an ATP-dependent conformational change of the enzyme, since inhibition only occurred when ATP was present during the antibody binding step. The 14-kDa subunit could be stripped from the membrane by treatment with the chaotropic agent KI, confirming it to be part of the soluble complex of the V-ATPase. In immunoblots, the 14-kDa-specific antibodies showed no cross-reaction with several xenic V-ATPases.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
4
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pubmed:volume |
269
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3767-74
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:8106421-Amino Acid Sequence,
pubmed-meshheading:8106421-Animals,
pubmed-meshheading:8106421-Antibodies, Monoclonal,
pubmed-meshheading:8106421-Base Sequence,
pubmed-meshheading:8106421-Blotting, Southern,
pubmed-meshheading:8106421-Blotting, Western,
pubmed-meshheading:8106421-Cloning, Molecular,
pubmed-meshheading:8106421-DNA, Complementary,
pubmed-meshheading:8106421-Digestive System,
pubmed-meshheading:8106421-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8106421-Gene Library,
pubmed-meshheading:8106421-Kinetics,
pubmed-meshheading:8106421-Larva,
pubmed-meshheading:8106421-Macromolecular Substances,
pubmed-meshheading:8106421-Molecular Sequence Data,
pubmed-meshheading:8106421-Moths,
pubmed-meshheading:8106421-Proton-Translocating ATPases,
pubmed-meshheading:8106421-Recombinant Fusion Proteins,
pubmed-meshheading:8106421-Restriction Mapping,
pubmed-meshheading:8106421-Vacuoles
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pubmed:year |
1994
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pubmed:articleTitle |
A novel 14-kDa V-ATPase subunit in the tobacco hornworm midgut.
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pubmed:affiliation |
Zoological Institute, University of Munich, Germany.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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