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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
1992-5-6
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pubmed:abstractText |
The outer membrane of Gram-negative bacteria presents an effective barrier that restricts the release of proteins from the cell. Virtually all extracellular proteins of Gram-negative bacteria are exported by specialized systems requiring the action of several gene products. We have constructed a tripartite fusion consisting of (i) the signal sequence and first nine N-terminal amino acids of the mature major Escherichia coli lipoprotein, (ii) amino acids 46-159 of the outer membrane protein OmpA, and (iii) the complete mature beta-lactamase (EC 3.5.2.6) sequence. This protein had an enzymatically active beta-lactamase and was found predominantly in the outer membrane. Immunofluorescence microscopy, the accessibility of the fusion protein to externally added proteases, and the rates of hydrolysis of nitrocefin and penicillin G by whole cells demonstrated that a substantial fraction (20-30%) of the beta-lactamase domain of the fusion protein was exposed on the external surface of E. coli. In cells grown at 24 degrees C the localization of beta-lactamase on the cell surface was almost quantitative (greater than 80% of the enzymatically active protein was exposed to the extracellular fluid) as determined by nitrocefin and penicillin G hydrolysis and trypsin accessibility. These results demonstrated that a soluble protein, beta-lactamase, can be transported through--and become anchored on--the outer membrane by fusion to the proper targeting and localization signals.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-1367632,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-1692799,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-1702781,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-1848301,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-1856199,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-1896445,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2124971,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2139651,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2181242,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2233249,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2247164,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2431904,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2468182,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2471451,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2543905,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2553664,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2684781,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2689448,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2853688,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-2997584,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3025450,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3047120,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3047121,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3283107,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3284654,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3330755,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3537309,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3902787,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-3912172,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-4555955,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-6336734,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-6368539,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-6425288,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-773686,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1557377-803320
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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 |
Apr
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
89
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2713-7
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pubmed:dateRevised |
2010-9-7
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pubmed:meshHeading |
pubmed-meshheading:1557377-Amino Acid Sequence,
pubmed-meshheading:1557377-Bacterial Outer Membrane Proteins,
pubmed-meshheading:1557377-Bacterial Proteins,
pubmed-meshheading:1557377-Biological Transport,
pubmed-meshheading:1557377-Cell Compartmentation,
pubmed-meshheading:1557377-Escherichia coli,
pubmed-meshheading:1557377-Genetic Engineering,
pubmed-meshheading:1557377-Lipoproteins,
pubmed-meshheading:1557377-Molecular Sequence Data,
pubmed-meshheading:1557377-Recombinant Fusion Proteins,
pubmed-meshheading:1557377-Structure-Activity Relationship,
pubmed-meshheading:1557377-beta-Lactamases
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pubmed:year |
1992
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pubmed:articleTitle |
Transport and anchoring of beta-lactamase to the external surface of Escherichia coli.
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pubmed:affiliation |
Department of Chemical Engineering, University of Texas, Austin 78712.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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