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Determination of substrate specificity and putative substrates of Chk2 kinase
Authors:Seo Gil-Ju  Kim Se-Eun  Lee Young-Man  Lee Jeong-Won  Lee Jae-Rin  Hahn Myong-Joon  Kim Seong-Tae
Institution:Department of Molecular Cellular Biology, Sungkyunkwan University School of Medicine, Samsung Biomedical Research Institute, 300 Chunchun-Dong, Changan-Ku, 440-746, Suwon, Kyunggi-Do, Republic of Korea.
Abstract:Chk2/hCds1, the human homolog of Saccharomyces cerevisiae Rad53p and Schizosaccharomyces pombe Cds1p, plays a critical role in the DNA damage checkpoint pathway. While several in vivo targets of Chk2 have been identified, the other target proteins of Chk2 responsible for multiple functions, such as cell cycle arrest, DNA repair, and apoptosis, remain to be elucidated. We utilized the GST-peptide approach to identify physiological substrates for Chk2. Mutational analyses using GST-linked Cdc25A containing serine 123 revealed that residues at positions -5 and -3 are critical determinants for the recognition of the Chk2 substrate. We determined the general phosphorylation consensus sequence and identified in vitro targets of Chk2 using GST peptides as substrates. The newly identified in vitro target proteins include Abl1, Bub1R, Bub1, Bub3, Psk-H1, Smc3, Plk1, Cdc25B, Dcamkl1, Mre11, Pms1, and Xrcc9.
Keywords:Chk2  Cdc25A  GST-peptide method  DNA damage response pathway  Substrate specificity
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