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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
1994-6-22
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pubmed:abstractText |
This paper describes a highly sensitive new method for the identification of penicillin-binding proteins (PBPs) that is based on the use of an ampicillin-digoxigenin conjugate (DIG-AMP conjugate) which is detected by immunoblotting and chemiluminescence. The sensitivity of chemiluminescence permitted X-ray film exposure times to be decreased to minutes, as opposed to the days or weeks which are requisite when conventionally radiolabeled beta-lactams are used. Coupling of ampicillin to digoxigenin yielded a product containing digoxigenin (detected by chemiluminescence) which also was inhibitory for Staphylococcus aureus and Escherichia coli. Unconjugated digoxigenin at concentrations of up to 100 micrograms/ml was not inhibitory for either organism. For S. aureus the MICs of DIG-AMP (0.7 microgram of conjugated ampicillin per ml) and of free ampicillin (0.5 microgram/ml) were comparable, indicating that ampicillin retained its bioactivity when coupled to digoxigenin. However, for E. coli the MICs of DIG-AMP (70 micrograms of conjugated ampicillin per ml) and of free ampicillin (8 micrograms/ml) were widely disparate, suggesting that the DIG-AMP conjugate was too large and/or hydrophobic to traverse the E. coli outer membrane via porins. DIG-AMP binding assays with E. coli and S. aureus cell envelopes revealed profiles of PBPs similar to those detected with 125I-ampicillin or [3H]penicillin. DIG-AMP binding to PBPs was completely inhibited in competition experiments with free ampicillin or penicillin, supporting the specificity of the DIG-AMP conjugate for PBPs. DIG-AMP thus represents an advantageous alternative to radioactive beta-lactams for the identification and analysis of PBPs.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-1091862,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-1103132,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-1370901,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-1640833,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-2076199,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-2076201,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-2647634,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-2670908,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-2677607,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-3055170,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-319999,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-344137,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-3531167,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-3850810,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-388439,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-4733239,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-6337993,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-6373702,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-6450748,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-6778384,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-6778388,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-7416741,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-8385929,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8192459-8438948
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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 |
0066-4804
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
38
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
330-6
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pubmed:dateRevised |
2011-11-17
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pubmed:meshHeading |
pubmed-meshheading:8192459-Ampicillin,
pubmed-meshheading:8192459-Bacterial Proteins,
pubmed-meshheading:8192459-Carrier Proteins,
pubmed-meshheading:8192459-Chromatography, High Pressure Liquid,
pubmed-meshheading:8192459-Digoxigenin,
pubmed-meshheading:8192459-Escherichia coli,
pubmed-meshheading:8192459-Hexosyltransferases,
pubmed-meshheading:8192459-Immunoblotting,
pubmed-meshheading:8192459-Luminescent Measurements,
pubmed-meshheading:8192459-Muramoylpentapeptide Carboxypeptidase,
pubmed-meshheading:8192459-Penicillin-Binding Proteins,
pubmed-meshheading:8192459-Peptidyl Transferases,
pubmed-meshheading:8192459-Sensitivity and Specificity
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pubmed:year |
1994
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pubmed:articleTitle |
Digoxigenin-ampicillin conjugate for detection of penicillin-binding proteins by chemiluminescence.
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pubmed:affiliation |
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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