Bacterial expression of an active tyrosine kinase from a protein A/truncated c-src fusion protein

Hideyuki Saya, Polly S.Y. Lee, Toru Nishi, Ichiro Izawa, Motowo Nakajima, Gary E. Gallick, Victor A. Levin

研究成果: Article

12 引用 (Scopus)

抜粋

The carboxy-terminal half of the c-src protein fused to the protein A moiety was expressed in bacteria. The protein A/truncated c-src fusion protein, which does not have SH2 and SH3 domains, is found in the periplasmic space allowing for a simple one-step purification and demonstrated high efficiency in autophosphorylation and exogeneous substrate phosphorylation. The missense mutation at codon 294 (Ile → Thr), which is located in the ATP-binding domain of the c-src, resulted in dramatic reduction of tyrosine kinase activity of the fusion protein. Using the fusion protein. we also revealed that staurosporin, a well-known kinase inhibitor, directly affects autophosphorylation of the C-terminal half of the c-src protein. This truncated c-src expression system provides a good source of enzyme for diverse experiments and is an ideal model for understanding the implication of structural alterations in the catalytic activity of the c-src kinase by site-directed mutagenesis experiments.

元の言語English
ページ(範囲)224-230
ページ数7
ジャーナルFEBS Letters
327
発行部数2
DOI
出版物ステータスPublished - 1993 7 26

    フィンガープリント

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

これを引用

Saya, H., Lee, P. S. Y., Nishi, T., Izawa, I., Nakajima, M., Gallick, G. E., & Levin, V. A. (1993). Bacterial expression of an active tyrosine kinase from a protein A/truncated c-src fusion protein. FEBS Letters, 327(2), 224-230. https://doi.org/10.1016/0014-5793(93)80174-S