Inhibition of a viral enzyme by a small-molecule dimer disruptor

Tina Shahian; Gregory M. Lee; Ana Lazic; Leggy A. Arnold; Priya Velusamy; Christina M. Roels; R. Kiplin Guy; Charles S. Craik

doi:10.1038/nchembio.192

Inhibition of a viral enzyme by a small-molecule dimer disruptor

Tina Shahian
, Gregory M. Lee
, Ana Lazic
, Leggy A. Arnold
, Priya Velusamy
, Christina M. Roels
, R. Kiplin Guy
, Charles S. Craik

Producción científica: Article › revisión exhaustiva

73 Citas (Scopus)

Resumen

We identified small-molecule dimer disruptors that inhibit an essential dimeric protease of human Kaposi's sarcoma-associated herpesvirus (KSHV) by screening an α-helical mimetic library. Next, we synthesized a second generation of low-micromolar inhibitors with improved potency and solubility. Complementary methods including size exclusion chromatography and 1H-13C HSQC titration using selectively labeled 13C-Met samples revealed that monomeric protease is enriched in the presence of inhibitor. 1H-15N HSQC titration studies mapped the inhibitor binding site to the dimer interface, and mutagenesis studies targeting this region were consistent with a mechanism where inhibitor binding prevents dimerization through the conformational selection of a dynamic intermediate. These results validate the interface of herpesvirus proteases and other similar oligomeric interactions as suitable targets for the development of small-molecule inhibitors.

Idioma original	English
Páginas (desde-hasta)	640-646
Número de páginas	7
Publicación	Nature Chemical Biology
Volumen	5
N.º	9
DOI	https://doi.org/10.1038/nchembio.192
Estado	Published - sept 2009

Nota bibliográfica

Funding Information:
The authors thank W. Gibson (The Johns Hopkins University School of Medicine) for providing us with the CMV protease expression plasmid and for valuable feedback on the manuscript. This work was funded by US National Institutes of Health grants T32 GMO7810 and AIO67423 (C.S.C.) and by the American Lebanese and Syrian Associated Charities and the St. Jude Children’s Research Hospital (R.K.G.). We also thank the University of California, San Francisco–Gladstone Institute for Virology and Immunology/Center for AIDS Research for the Clinical Science Pilot Award (P30-AI027763 to G.M.L.).

Financiación

The authors thank W. Gibson (The Johns Hopkins University School of Medicine) for providing us with the CMV protease expression plasmid and for valuable feedback on the manuscript. This work was funded by US National Institutes of Health grants T32 GMO7810 and AIO67423 (C.S.C.) and by the American Lebanese and Syrian Associated Charities and the St. Jude Children’s Research Hospital (R.K.G.). We also thank the University of California, San Francisco–Gladstone Institute for Virology and Immunology/Center for AIDS Research for the Clinical Science Pilot Award (P30-AI027763 to G.M.L.).

Financiadores	Número del financiador
National Institutes of Health (NIH)	AIO67423, T32 GMO7810
National Institute of Allergy and Infectious Diseases	R01AI067423
St. Jude Children's Research Hospital
American Lebanese Syrian Associated Charities

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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title = "Inhibition of a viral enzyme by a small-molecule dimer disruptor",

abstract = "We identified small-molecule dimer disruptors that inhibit an essential dimeric protease of human Kaposi's sarcoma-associated herpesvirus (KSHV) by screening an α-helical mimetic library. Next, we synthesized a second generation of low-micromolar inhibitors with improved potency and solubility. Complementary methods including size exclusion chromatography and 1H-13C HSQC titration using selectively labeled 13C-Met samples revealed that monomeric protease is enriched in the presence of inhibitor. 1H-15N HSQC titration studies mapped the inhibitor binding site to the dimer interface, and mutagenesis studies targeting this region were consistent with a mechanism where inhibitor binding prevents dimerization through the conformational selection of a dynamic intermediate. These results validate the interface of herpesvirus proteases and other similar oligomeric interactions as suitable targets for the development of small-molecule inhibitors.",

author = "Tina Shahian and Lee, \{Gregory M.\} and Ana Lazic and Arnold, \{Leggy A.\} and Priya Velusamy and Roels, \{Christina M.\} and Guy, \{R. Kiplin\} and Craik, \{Charles S.\}",

note = "Funding Information: The authors thank W. Gibson (The Johns Hopkins University School of Medicine) for providing us with the CMV protease expression plasmid and for valuable feedback on the manuscript. This work was funded by US National Institutes of Health grants T32 GMO7810 and AIO67423 (C.S.C.) and by the American Lebanese and Syrian Associated Charities and the St. Jude Children{\textquoteright}s Research Hospital (R.K.G.). We also thank the University of California, San Francisco–Gladstone Institute for Virology and Immunology/Center for AIDS Research for the Clinical Science Pilot Award (P30-AI027763 to G.M.L.).",

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AU - Roels, Christina M.

AU - Guy, R. Kiplin

AU - Craik, Charles S.

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PY - 2009/9

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AB - We identified small-molecule dimer disruptors that inhibit an essential dimeric protease of human Kaposi's sarcoma-associated herpesvirus (KSHV) by screening an α-helical mimetic library. Next, we synthesized a second generation of low-micromolar inhibitors with improved potency and solubility. Complementary methods including size exclusion chromatography and 1H-13C HSQC titration using selectively labeled 13C-Met samples revealed that monomeric protease is enriched in the presence of inhibitor. 1H-15N HSQC titration studies mapped the inhibitor binding site to the dimer interface, and mutagenesis studies targeting this region were consistent with a mechanism where inhibitor binding prevents dimerization through the conformational selection of a dynamic intermediate. These results validate the interface of herpesvirus proteases and other similar oligomeric interactions as suitable targets for the development of small-molecule inhibitors.

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Inhibition of a viral enzyme by a small-molecule dimer disruptor

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Nota bibliográfica

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ASJC Scopus subject areas

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