Conversion of a Tyrosine Kinase Protein Substrate to a High Affinity Ligand by ATP Linkage

Kui Shen, Philip A. Cole

Research output: Contribution to journalArticle

Abstract

Protein kinases often show low affinity for their protein substrates, which makes it difficult to study kinase-substrate interactions. Here, we show using expressed protein ligation with the signaling protein Src that it is feasible to install a covalently linked ATP moiety into the tail of Src, generating a semisynthetic protein with a high affinity for its cognate tyrosine kinase, Csk. It is also established that this Src-ATP conjugate can be used to selectively pull down Csk from a complex protein mixture. This work outlines a general strategy for identifying an unknown kinase that is responsible for the phosphorylation of a protein substrate on a site of interest.

Original languageEnglish (US)
Pages (from-to)16172-16173
Number of pages2
JournalJournal of the American Chemical Society
Volume125
Issue number52
DOIs
StatePublished - Dec 31 2003

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ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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