Identification of potential binding sites for the FHA domain of human Chk2 by in vitro binding studies.
Human Chk2 is a newly identified tumor suppressor protein involved in signaling pathways in response to DNA damage. The protein consists of a forkhead-associated (FHA) domain and a kinase domain. Identification of binding partners of the Chk2FHA domain is important in understanding the roles of Chk2 in signaling. We report development of an approach involving the use of combinatorial libraries, pull-down assays, surface plasmon resonance (SPR), and nuclear magnetic resonance (NMR) methods to identify possible candidates for the binding sites of Chk2FHA. The approach has been used to identify Thr329 of p53 and Thr1852 of breast cancer type 1 susceptibility protein (BRCA1) as very likely biological binding sites of Chk2FHA. The results provide useful leads for further biological analyses of cell signaling involving the FHA domain of Chk2 protein.
Qin
Dongyan
D
The Ohio State Biochemistry Program, The Ohio State University, Columbus, OH 43220, USA.
Lee
Hyun
H
Yuan
Chunhua
C
Ju
Yong
Y
Tsai
Ming-Daw
MD
D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry*; D064447:Checkpoint Kinase 2; D051858:Forkhead Transcription Factors; D006801:Humans; D009687:Nuclear Proteins / Q000737:chemistry*; D019151:Peptide Library; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000145:classification; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000737:chemistry*; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry*; D044767:Ubiquitin-Protein Ligases
D002352:Carrier Proteins; D051858:Forkhead Transcription Factors; D009687:Nuclear Proteins; D019151:Peptide Library; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; C111164:BRAP protein, human; D044767:Ubiquitin-Protein Ligases; D064447:Checkpoint Kinase 2; C578634:CHEK2 protein, human; D017346:Protein Serine-Threonine Kinases