Abstract
Kinases have become popular therapeutic targets primarily due to their integral role in cell cycle and tumor progression. The efficacy of high-throughput screening efforts is dependent on the development of high quality multiplex tools capable of replacing lower-throughput technologies such as mass spectroscopy or solution-based assays for the study of kinase–substrate interactions. Functional protein microarrays are comprised of thousands of immobilized proteins on glass slides that have been used successfully to identify protein–protein interactions. Here, we describe the application of functional protein microarrays for the identification of the phosphorylation targets of individual protein kinases using highly sensitive radioactive detection and robust informatics algorithms.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Gershon D (2003) Proteomics technologies: probing the proteome. Nature 424:581–587
MacBeath G, Schreiber SL (2000) Printing proteins as microarrays for high-throughput function determination. Science 289:1760–1763
Phizicky E, Bastiaens PI, Zhu H, Snyder M, Fields S (2003) Protein analysis on a proteomic scale. Nature 422:208–215
Zhu H, Bilgin M, Bangham R, Hall D, Casamayor A, Bertone P, Lan N, Jansen R, Bidlingmaier S, Houfek T, Mitchell T, Miller P, Dean RA, Gerstein M, Snyder M (2001) Global analysis of protein activities using proteome chips. Science 293:2101–2105
Ptacek J, Devgan G, Michaud G, Zhu H, Zhu X, Fasolo J, Guo H, Jona G, Breitkreutz A, Sopko R, McCartney RR, Schmidt MC, Rachidi N, Lee SJ, Mah AS, Meng L, Stark MJ, Stern DF, De Virgilio C, Tyers M, Andrews B, Gerstein M, Schweitzer B, Predki PF, Snyder M (2005) Global analysis of protein phosphorylation in yeast. Nature 438:679–684
Gelperin DM, White MA, Wilkinson ML, Kon Y, Kung LA, Wise KJ, Lopez-Hoyo N, Jiang L, Piccirillo S, Yu H, Gerstein M, Dumont ME, Phizicky EM, Snyder M, Grayhack EJ (2005) Biochemical and genetic analysis of the yeast proteome with a movable ORF collection. Genes Dev 19:2816–2826
Schweitzer B, Predki P, Snyder M (2003) Microarrays to characterize protein interactions on a whole-proteome scale. Proteomics 11:2190–2199
Haab B (2003) Methods and applications of antibody microarrays in cancer research. Proteomics 11:2116–2122
Manning G, Whyte DB, Martiniez R, Hunter T, Sudarsanam S (2002) The protein kinase complement of the human genome. Science 298:1912–1934
Zhu X, Gerstein M, Snyder M (2006) ProCAT: a data analysis approach for protein microarrays. Genome Biol 7:R110
Zhang JH, Chung TD, Oldenburg KR (2000) Confirmation of primary active substances from high throughput screening of chemical and biological populations: a statistical approach and practical considerations. J Comb Chem 3:258–265
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Smith, M.G., Ptacek, J., Snyder, M. (2011). Kinase Substrate Interactions. In: Wu, C. (eds) Protein Microarray for Disease Analysis. Methods in Molecular Biology, vol 723. Humana Press. https://doi.org/10.1007/978-1-61779-043-0_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-043-0_13
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-042-3
Online ISBN: 978-1-61779-043-0
eBook Packages: Springer Protocols