Abstract
The promise of rapid and cost-effective drug screening assays on solid support is one that may now be realized with the advent of small molecule microarrays. Many of the initial hurdles in library design and microarray fabrication have been overcome over the last decade, allowing this platform to become more accessible to researchers across both the academic and industrial spheres. Beyond pharmaceutical screening, microarrays reveal quantitative ligand-binding signatures that in the form of protein fingerprints provide a means to discriminate between closely related proteins. The value of protein fingerprinting in drug discovery is also highlighted through the identification of ligands that not only offer good potency, but also good selectivity. Herein, we describe the method for high-throughput screening and profiling of metalloproteases on small molecule microarrays. Metalloproteases are an important class of proteins, which are implicated in the pathogenicity of certain microbes and in the progression of cancer. We have introduced a novel two-colour labelling and application approach that directly elucidates functional ligands, reducing the burden of downstream revalidation of identified hits.
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(2007) The academic pursuit of screening. Nat Chem Biol 3:433
Uttamchandani M, Walsh DP, Yao SQ, Chang YT (2005) Small molecule microarrays: recent advances and applications. Curr Opin Chem Biol 9:4-13
Hu Y, Uttamchandani M, Yao SQ (2006) Microarray: a versatile platform for high-throughput functional proteomics. Comb Chem High Throughput Screen 9:203-212
Duffner JL, Clemons PA, Koehler AN (2007) A pipeline for ligand discovery using small-molecule microarrays. Curr Opin Chem Biol 11:74-82
Uttamchandani M, Wang J, Yao SQ (2006) Protein and small molecule microarrays: powerful tools for high-throughput proteomics. Mol Biosyst 2:58-68
Uttamchandani M, Lee WL, Wang J, Yao SQ (2007) Quantitative inhibitor fingerprinting of metalloproteases using small molecule microarrays. J Am Chem Soc 129:13110-13117
Lee WL, Li J, Uttamchandani M, Sun H, Yao SQ (2007) Inhibitor fingerprinting of metalloproteases using microplate and microarray platforms: an enabling technology in Catalomics. Nat Protoc 2:2126-2138
Wang J, Uttamchandani M, Sun LP and Yao SQ (2006) Activity-based high-throughput profiling of metalloprotease inhibitors using small molecule microarrays. Chem Commun (Camb) 717-719
Wang J, Uttamchandani M, Li J, Hu M, Yao SQ (2006) Rapid assembly of matrix metalloprotease inhibitors using click chemistry. Org Lett 8:3821-3824
Uttamchandani M, Wang J, Li J, Hu M, Sun H, Chen KY, Liu K, Yao SQ (2007) Inhibitor fingerprinting of matrix metalloproteases using a combinatorial peptide hydroxamate library. J Am Chem Soc 129:7848-7858
Chan WC, White PD (2000) Fmoc solid phase peptide synthesis. Oxford University Press, New York
Acknowledgments
Funding support is acknowledged from DSO National Laboratories and the National University of Singapore.
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Uttamchandani, M. (2010). High-Throughput Screening of Metalloproteases Using Small Molecule Microarrays. In: Chittur, S.V. (eds) Microarray Methods for Drug Discovery. Methods in Molecular Biology, vol 632. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-663-4_13
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DOI: https://doi.org/10.1007/978-1-60761-663-4_13
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Publisher Name: Humana Press, Totowa, NJ
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