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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
6 Pt 1
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pubmed:dateCreated |
1996-3-7
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pubmed:abstractText |
Protein metabolic labeling in vivo was used to determine a time course for trafficking of nascent H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase) from endoplasmic reticulum (ER) to mature tubulovesicles in parietal cells. Stomachs of cimetidine-treated rabbits were taken 15-90 min after injection of [35S]methionine/cysteine, and mucosal microsomes were fractionated on sucrose gradients for analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot, and autoradiography. After 15 min, labeled alpha-subunit peaked at approximately 1.14 g/ml, matching the distribution of the high-mannose beta-subunit precursor, "pre-beta." After 30 min, most labeled alpha-subunit was in a peak at approximately 1.10 g/ml, considered to be Golgi. By 90 min, most labeled alpha-subunit was in a light peak, at approximately 1.07 g/ml, aligned with the major peak of total H(+)-K(+)-ATPase previously characterized as mature tubulovesicles. From material enriched in pre-beta, alpha-subunit was coprecipitated with pre-beta by a terminal mannose-specific lectin, Galanthus nivalis agglutinin, in the same ratio as the mature alpha:beta ratio. Thus alpha- and beta-subunits associated early in the ER. This is the first use of protein metabolic labeling to study early trafficking of the H(+)-K(+)-ATPase in vivo. The techniques may be usefully applied to examining changes in H(+)-K(+)-ATPase synthetic rate in response to various pharmacological treatments and studying the divergent pathways for nascent H(+)-K(+)- and Na(+)-K(+)-ATPases.
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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 |
Dec
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pubmed:issn |
0002-9513
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pubmed:author | |
pubmed:issnType |
Print
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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 |
G883-91
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:8572220-Animals,
pubmed-meshheading:8572220-Centrifugation, Density Gradient,
pubmed-meshheading:8572220-Chromatography, Affinity,
pubmed-meshheading:8572220-Detergents,
pubmed-meshheading:8572220-Drug Resistance,
pubmed-meshheading:8572220-Endoplasmic Reticulum,
pubmed-meshheading:8572220-Galanthus,
pubmed-meshheading:8572220-H(+)-K(+)-Exchanging ATPase,
pubmed-meshheading:8572220-Microsomes,
pubmed-meshheading:8572220-Parietal Cells, Gastric,
pubmed-meshheading:8572220-Rabbits,
pubmed-meshheading:8572220-Subcellular Fractions,
pubmed-meshheading:8572220-Time Factors
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pubmed:year |
1995
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pubmed:articleTitle |
In vivo trafficking of nascent H(+)-K(+)-ATPase in rabbit parietal cells.
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pubmed:affiliation |
Department of Molecular and Cell Biology, University of California, Berkeley 94720-3200, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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