Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors.

Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Research Abstract Details 

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Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Abstract Text:

Hyun-Ku Lee,Steven J Brown,Hugh Rosen,Peter S Tobias,

We describe a successful application of beta-lactamase fragment complementation to high-throughput screening (HTS) for Toll-like receptor 4 (TLR4) signaling inhibitors. We developed a stable cell line, HeLa/CL3-4, expressing MyD88/Bla(a) and TLR4/Bla(b), in which the two beta-lactamase fragments complement with each other by virtue of spontaneous MyD88-TLR4 binding via their Toll/IL-1R (TIR) domains. Inhibition of the MyD88-TLR4 binding leads to the disruption of the enzyme complementation and a loss of the lactamase activity. We used a 384-well plate format to screen 16,000 compounds using this assay and obtained 45 primary hits. After rescreening these 45 hits and eliminating compounds that directly inhibited beta-lactamase, we had five candidate inhibitors. We show that these five act as inhibitors of TLR4-MyD88 binding and are variously effective at inhibiting lipopolysaccharide-stimulated cytokine release from RAW264.7 cells. One compound is effective near 100 nM. None of the compounds showed any cytotoxicity at 20 microM.

Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Publishing Authors By Initials

HK Lee,SJ Brown,H Rosen,PS Tobias,

For similar enzymes and coenzymes: enzymes: hydrolases: amidohydrolases: beta-lactamases research abstracts see: enzymes and coenzymes: enzymes: hydrolases: amidohydrolases: beta-lactamases research

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Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Molecular pharmacology

VOLUME: 72

Page Numbers: 868-75

Journal Abbreviation: Mol. Pharmacol.

ISSN: 0026-895X

DAY: 5

MONTH: 07

YEAR: 2007

Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 35623

Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Keywords Mesh Terms:

KEYWORDS: beta-Lactamases

MESH TERMS: metabolism

Chemical & Substance for Abstract: Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors. Information

Substance Name: beta-Lactamases

Registry Number: EC 3.5.2.6

Grant and Affiliation Information for Application of beta-lactamase enzyme complementation to the high-throughput screening of toll-like receptor signaling inhibitors.

AFFILIATION: Department of Immunology, IMM-12, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. tobias@scripps.edu

Country: United States

AGENCY: United States NIMH

GRANT: MH081265

ACRONYM: MH

MEDLINETA: Mol Pharmacol

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