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A rapid method for determining protein kinase phosphorylation specificity

Nature Methods volume 1, pages 27–29 (2004)Cite this article

Abstract

Selection of target substrates by protein kinases is strongly influenced by the amino acid sequence surrounding the phosphoacceptor site. Identification of the preferred peptide phosphorylation motif for a given kinase permits the production of efficient peptide substrates and greatly simplifies the mapping of phosphorylation sites in protein substrates. Here we describe a combinatorial peptide library method that allows rapid generation of phosphorylation motifs for serine/threonine kinases.

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Figure 1: Determination of phosphorylation motifs for serine/threonine kinases.
Figure 2: Pim2 phosphorylation specificity.

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Acknowledgements

We thank N. Ahn for providing the active MKK1 clone and T. Obata for preparation of recombinant Akt used in pilot studies. This work was supported by US National Institutes Health grants GM56203 to L.C.C. and CA75134 to A.T. B.E.T. is a Leukemia and Lymphoma Society Special Fellow.

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Authors and Affiliations

  1. Division of Signal Transduction, Harvard Medical School, 330 Brookline Avenue, Boston, 02215, Massachusetts, USA

    Jessica E Hutti, Emily T Jarrell, James D Chang, Derek W Abbott, Lewis C Cantley & Benjamin E Turk

  2. Division of Cardiology, Harvard Medical School, 330 Brookline Avenue, Boston, 02215, Massachusetts, USA

    James D Chang

  3. Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, 02215, Massachusetts, USA

    Peter Storz & Alex Toker

  4. Department of Systems Biology, Harvard Medical School, 330 Brookline Avenue, Boston, 02215, Massachusetts, USA

    Jessica E Hutti, Emily T Jarrell, Derek W Abbott, Lewis C Cantley & Benjamin E Turk

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  1. Jessica E Hutti
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Correspondence to Benjamin E Turk.

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The authors declare no competing financial interests.

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Hutti, J., Jarrell, E., Chang, J. et al. A rapid method for determining protein kinase phosphorylation specificity. Nat Methods 1, 27–29 (2004). https://doi.org/10.1038/nmeth708

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