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Peptide Microarrays pp 27–34Cite as

Analysis of Protein Tyrosine Kinase Specificity Using Positional Scanning Peptide Microarrays

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1352))

Abstract

Protein tyrosine kinases phosphorylate their substrates within the context of specific consensus sequences surrounding the site of modification. We describe a peptide microarray approach to rapidly determine tyrosine kinase phosphorylation site motifs. This method uses a peptide library that systematically substitutes each of the amino acid residues at multiple positions surrounding a central tyrosine residue. Peptide substrates are synthesized as biotin conjugates for immobilization on avidin-coated slides. Following incubation of the slide with protein kinase and radiolabeled ATP, the relative extent of phosphorylation of each of the peptides is quantified by phosphor imaging. This method allows small quantities of kinase to be analyzed rapidly in parallel, facilitating analysis of large numbers of kinases.

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Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, Yale University, New Haven, CT, USA

    Yang Deng

  2. Department of Pharmacology, Yale University School of Medicine, SHM B395e, 333 Cedar Street, New Haven, CT, 06520, USA

    Benjamin E. Turk

Authors
  1. Yang Deng
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  2. Benjamin E. Turk
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Corresponding author

Correspondence to Benjamin E. Turk .

Editor information

Editors and Affiliations

  1. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marina Cretich

  2. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marcella Chiari

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Deng, Y., Turk, B.E. (2016). Analysis of Protein Tyrosine Kinase Specificity Using Positional Scanning Peptide Microarrays. In: Cretich, M., Chiari, M. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 1352. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3037-1_3

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  • DOI: https://doi.org/10.1007/978-1-4939-3037-1_3

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3036-4

  • Online ISBN: 978-1-4939-3037-1

  • eBook Packages: Springer Protocols

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