BOCILLIN FL, a Sensitive and Commercially Available Reagent for Detection of Penicillin-Binding Proteins
| Autor: | Steven D. Kahl, Larry C. Blaszczak, Genshi Zhao, Kyle R. Gee, Timothy I. Meier |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Boron Compounds
Penicillin binding proteins medicine.drug_class Antibiotics Penicillins Muramoylpentapeptide Carboxypeptidase Sensitivity and Specificity Bacterial cell structure Microbiology chemistry.chemical_compound Bacterial Proteins polycyclic compounds medicine Penicillin-Binding Proteins Pharmacology (medical) Mechanisms of Action: Physiological Effects Fluorescent Dyes Pharmacology chemistry.chemical_classification Gel electrophoresis Bacteria biology biology.organism_classification Penicillin Infectious Diseases Enzyme Hexosyltransferases Biochemistry chemistry Peptidyl Transferases Biological Assay Peptidoglycan Carrier Proteins Protein Binding medicine.drug |
| Zdroj: | Antimicrobial Agents and Chemotherapy. 43:1124-1128 |
| ISSN: | 1098-6596 0066-4804 |
| DOI: | 10.1128/aac.43.5.1124 |
| Popis: | Penicillin-binding proteins (PBPs) are the enzymes that are required for the biosynthesis of the bacterial cell wall (10, 11, 23, 31). PBPs catalyze the final steps of the polymerization (transglycosylation) and cross-linking (transpeptidation) of peptidoglycan, an essential component of the bacterial cell wall (10, 11, 23, 31). PBPs are membrane-bound enzymes and targets of β-lactam antibiotics (6, 7, 10, 11, 14, 24, 28–31). The emerging resistance of pathogenic gram-positive bacteria to β-lactam antibiotics is a serious clinical problem (6, 25, 29, 30). Resistance to β-lactam antibiotics in some species such as Streptococcus pneumoniae has occurred by development of altered high-molecular-mass PBPs which have reduced affinity for the antibiotics (6, 7, 10, 11, 14, 24, 25, 29, 30). Extensive molecular and genetic studies have been devoted to the understanding of the mechanisms of bacterial resistance to β-lactam antibiotics (6, 7, 10, 11, 25, 29, 30). PBPs have often been detected by labeling bacterial membrane preparations with 3H-, 14C-, or 125I-labeled penicillin, separating the labeled proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and exposing the gels to X-ray films (22, 27, 28). The major limitations of this type of methodology are time intensiveness (days to weeks), accumulation of hazardous materials, and lack of commercial availability. In this study, we describe a new, rapid, sensitive, and nonradioactive method for the detection and study of bacterial PBPs. This method involves the use of BOCILLIN FL (Fig. (Fig.1),1), a newly synthesized and commercially available fluorescent penicillin, as a labeling reagent (Molecular Probes, Inc., Eugene, Oreg.). The BOCILLIN FL-labeled PBPs were separated by SDS-PAGE and detected with a FluorImager or with the naked eye under UV light. FIG. 1 Structure of BOCILLIN FL. |
| Databáze: | OpenAIRE |
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