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- Publisher Website: 10.1021/ac102595r
- Scopus: eid_2-s2.0-79954596002
- PMID: 21338058
- WOS: WOS:000288182900020
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Article: Engineered Amp C β-lactamase as a fluorescent screening tool for class C β-lactamase inhibitors
Title | Engineered Amp C β-lactamase as a fluorescent screening tool for class C β-lactamase inhibitors |
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Authors | |
Issue Date | 2011 |
Publisher | American Chemical Society. The Journal's web site is located at http://pubs.acs.org/ac |
Citation | Analytical Chemistry, 2011, v. 83 n. 6, p. 1996-2004 How to Cite? |
Abstract | Class C β-lactamases mediate antibiotic resistance in bacteria by efficiently hydrolyzing a broad range of β-lactam antibiotics. With their clinical significance and the lack of commercially available effective inhibitors, development of class C β-lactamase inhibitors has become one of the recent hot issues in the pharmaceutical industry. In this paper, we report the protein engineering of a fluorescent Amp C β-lactamase mutant designated as V211Cf for the in vitro screening of class C β-lactamase inhibitors. When a fluorescein (f) was incorporated at the entrance of the enzyme's active site (position 211), Amp C β-lactamase from Enterobacter cloacae P99 was tailor-made into a novel fluorescent biosensing protein that could display a fluorescence enhancement upon binding with its β-lactam substrates/inhibitors. With its catalytic activity close to the wild-type level, V211Cf can act as a "natural" fluorescent drug target for screening small binding molecules. In addition, V211Cf can allow specific detection for its active-site binding molecules and discriminate them from nondruglike molecules in the screen. Furthermore, V211Cf is amenable to a high throughput format. Taken together, V211Cf demonstrates the potential as an efficient tool for screening class C β-lactamase inhibitors and facilitates the discovery of therapeutics that can combat the clinically important class C β-lactamases. © 2011 American Chemical Society. |
Persistent Identifier | http://hdl.handle.net/10722/168521 |
ISSN | 2023 Impact Factor: 6.7 2023 SCImago Journal Rankings: 1.621 |
ISI Accession Number ID | |
References |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Tsang, MW | en_US |
dc.contributor.author | Chan, PH | en_US |
dc.contributor.author | So, PK | en_US |
dc.contributor.author | Ma, DL | en_US |
dc.contributor.author | Tsang, CW | en_US |
dc.contributor.author | Wong, KY | en_US |
dc.contributor.author | Leung, YC | en_US |
dc.date.accessioned | 2012-10-08T03:19:58Z | - |
dc.date.available | 2012-10-08T03:19:58Z | - |
dc.date.issued | 2011 | en_US |
dc.identifier.citation | Analytical Chemistry, 2011, v. 83 n. 6, p. 1996-2004 | en_US |
dc.identifier.issn | 0003-2700 | en_US |
dc.identifier.uri | http://hdl.handle.net/10722/168521 | - |
dc.description.abstract | Class C β-lactamases mediate antibiotic resistance in bacteria by efficiently hydrolyzing a broad range of β-lactam antibiotics. With their clinical significance and the lack of commercially available effective inhibitors, development of class C β-lactamase inhibitors has become one of the recent hot issues in the pharmaceutical industry. In this paper, we report the protein engineering of a fluorescent Amp C β-lactamase mutant designated as V211Cf for the in vitro screening of class C β-lactamase inhibitors. When a fluorescein (f) was incorporated at the entrance of the enzyme's active site (position 211), Amp C β-lactamase from Enterobacter cloacae P99 was tailor-made into a novel fluorescent biosensing protein that could display a fluorescence enhancement upon binding with its β-lactam substrates/inhibitors. With its catalytic activity close to the wild-type level, V211Cf can act as a "natural" fluorescent drug target for screening small binding molecules. In addition, V211Cf can allow specific detection for its active-site binding molecules and discriminate them from nondruglike molecules in the screen. Furthermore, V211Cf is amenable to a high throughput format. Taken together, V211Cf demonstrates the potential as an efficient tool for screening class C β-lactamase inhibitors and facilitates the discovery of therapeutics that can combat the clinically important class C β-lactamases. © 2011 American Chemical Society. | en_US |
dc.language | eng | en_US |
dc.publisher | American Chemical Society. The Journal's web site is located at http://pubs.acs.org/ac | en_US |
dc.relation.ispartof | Analytical Chemistry | en_US |
dc.subject.mesh | Anti-Bacterial Agents - Metabolism - Pharmacology | en_US |
dc.subject.mesh | Bacterial Proteins - Antagonists & Inhibitors - Chemistry - Genetics - Metabolism | en_US |
dc.subject.mesh | Biocatalysis | en_US |
dc.subject.mesh | Drug Evaluation, Preclinical - Methods | en_US |
dc.subject.mesh | Enterobacter Cloacae - Enzymology | en_US |
dc.subject.mesh | Enzyme Inhibitors - Metabolism - Pharmacology | en_US |
dc.subject.mesh | Models, Molecular | en_US |
dc.subject.mesh | Mutation | en_US |
dc.subject.mesh | Protein Conformation | en_US |
dc.subject.mesh | Protein Engineering | en_US |
dc.subject.mesh | Spectrometry, Fluorescence | en_US |
dc.subject.mesh | Beta-Lactamases - Antagonists & Inhibitors - Chemistry - Genetics - Metabolism | en_US |
dc.subject.mesh | Beta-Lactams - Metabolism - Pharmacology | en_US |
dc.title | Engineered Amp C β-lactamase as a fluorescent screening tool for class C β-lactamase inhibitors | en_US |
dc.type | Article | en_US |
dc.identifier.email | Ma, DL:edmondma@hku.hk | en_US |
dc.identifier.authority | Ma, DL=rp00760 | en_US |
dc.description.nature | link_to_subscribed_fulltext | en_US |
dc.identifier.doi | 10.1021/ac102595r | en_US |
dc.identifier.pmid | 21338058 | - |
dc.identifier.scopus | eid_2-s2.0-79954596002 | en_US |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-79954596002&selection=ref&src=s&origin=recordpage | en_US |
dc.identifier.volume | 83 | en_US |
dc.identifier.issue | 6 | en_US |
dc.identifier.spage | 1996 | en_US |
dc.identifier.epage | 2004 | en_US |
dc.identifier.isi | WOS:000288182900020 | - |
dc.publisher.place | United States | en_US |
dc.identifier.scopusauthorid | Tsang, MW=36720156000 | en_US |
dc.identifier.scopusauthorid | Chan, PH=36545749700 | en_US |
dc.identifier.scopusauthorid | So, PK=14919747700 | en_US |
dc.identifier.scopusauthorid | Ma, DL=7402075538 | en_US |
dc.identifier.scopusauthorid | Tsang, CW=7202935952 | en_US |
dc.identifier.scopusauthorid | Wong, KY=26641515400 | en_US |
dc.identifier.scopusauthorid | Leung, YC=35074432700 | en_US |
dc.identifier.issnl | 0003-2700 | - |