Expression of Human Tyrosine Kinase, Lck, in Yeast Saccharomyces cerevisiae: Growth Suppression and Strategy for Inhibitor Screening

Title: Expression of Human Tyrosine Kinase, Lck, in Yeast Saccharomyces cerevisiae: Growth Suppression and Strategy for Inhibitor Screening

Volume: 13 Issue: 9

Author(s): Masashi Koyama, Shoko Saito, Rumiko Nakagawa, Iwao Katsuyama, Masakazu Hatanaka, Tadashi Yamamoto, Tsutomu Arakawa and Masao Tokunaga

Affiliation:

Keywords: Tyrosine kinase, Lck, Expression, Saccharomyces cerevisiae, Tob, Inhibitor screening

Abstract: We report the successful expression and detection of a phosphorylated form of human T cell tyrosine kinase, Lck, in Saccharomyes cerevisiae, which leads to growth suppression of the yeast cells. Expression of an inactive Lck mutant resulted in no phosphorylation and no growth suppression, indicating that cell growth inhibition by Lck is due to the activity of the kinase, consistent with the observed tyrosine-phosphorylation of the Lck and yeast host cell proteins. The addition of a known inhibitor of Lck to the cell culture resulted in recovery of cell growth expressing the active Lck, suggesting that the growth inhibition by lck gene expression can be used to screen inhibitors for the gene product. We have extended such approach to Tob, another potential therapeutic target.

Cite this article as:

Koyama Masashi, Saito Shoko, Nakagawa Rumiko, Katsuyama Iwao, Hatanaka Masakazu, Yamamoto Tadashi, Arakawa Tsutomu and Tokunaga Masao, Expression of Human Tyrosine Kinase, Lck, in Yeast Saccharomyces cerevisiae: Growth Suppression and Strategy for Inhibitor Screening, Protein & Peptide Letters 2006; 13 (9) . https://dx.doi.org/10.2174/092986606778256216