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Microarray platform for profiling enzyme activities in complex proteomes

Academic Article
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Overview

authors

  • Sieber, S. A.
  • Mondala, T. S.
  • Head, Steve
  • Cravatt, Benjamin

publication date

  • December 2004

journal

  • Journal of the American Chemical Society  Journal

abstract

  • Activity-based protein profiling (ABPP) is a chemical method that utilizes active-site-directed probes to determine the functional state of enzymes in complex proteomes. Probe-labeled enzymes are typically detected by in-gel fluorescence scanning, a robust technique that nonetheless exhibits some key deficiencies, including limited sensitivity and resolution, as well as ambiguity regarding the molecular identity of the enzymes under investigation. Here, we report a microarray platform for ABPP that addresses these limitations. In this platform, proteomes are treated with ABPP probes in solution, after which labeled enzymes are captured and visualized on glass slides displaying an array of anti-enzyme antibodies. We show that ABPP microarrays exhibit superior sensitivity and resolution compared to gel-based methods, permitting the parallel analysis of several enzyme activities in proteomes, including cancer-associated proteases such as urokinase, matrix metalloproteinase-9, and prostate-specific antigen.

subject areas

  • Antibodies
  • Benzophenones
  • Binding Sites
  • Matrix Metalloproteinase 9
  • Protein Array Analysis
  • Proteome
  • Rhodamines
  • Sensitivity and Specificity
  • Serine Endopeptidases
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Identity

International Standard Serial Number (ISSN)

  • 0002-7863

Digital Object Identifier (DOI)

  • 10.1021/ja044286+

PubMed ID

  • 15571375
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Additional Document Info

start page

  • 15640

end page

  • 15641

volume

  • 126

issue

  • 48

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