title: High-Throughput Screening of Substrate Specificity for Protein Tyrosine Phosphatases (PTPs) on Phosphopeptide Microarrays.
abstract: Phosphatases are a family of enzymes responsible for the dephosphorylation of biomolecules. Phosphatases play essential roles in cell cycle regulation, signal transduction, and cellular communication. In recent years, one type of phosphatases, protein tyrosine phosphatases (PTPs), emerges as important therapeutic targets for complex and devastating diseases. Nevertheless, the physiological roles, substrate specificity, and downstream targets for PTPs remain largely unknown. To demonstrate how microarrays can be applied to characterizing PTPs, we describe here a phosphopeptide microarray strategy for activity-based high-throughput screening of PTPs substrate specificity. This is followed by a kinetic microarray assay and microplate assay to determine the rate constants of dephosphorylation by PTPs. This microarray strategy has been successfully applied to identifying several potent and selective substrates against different PTPs. These substrates could be used to design potent and selective PTPs inhibitors in the future.
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, Singapore, 138669, Singapore.|Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.
authors.forename: Liqian
authors.initials: L
authors.lastname: Gao
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, Singapore, 138669, Singapore.
authors.forename: Su Seong
authors.initials: SS
authors.lastname: Lee
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Key Lab of Nuclear Radiation and Nuclear Energy Technology, Center for Multidisciplinary Research, Chinese Academy of Sciences (CAS), Institute of High Energy Physics, 19B Yuquan Road, Beijing, 100049, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Biology and Chemistry, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong, P.R. China. hongysun@cityu.edu.hk.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Sun
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Key Lab of Nuclear Radiation and Nuclear Energy Technology, Center for Multidisciplinary Research, Chinese Academy of Sciences (CAS), Institute of High Energy Physics, 19B Yuquan Road, Beijing, 100049, China.
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Key Lab of Nuclear Radiation and Nuclear Energy Technology, Center for Multidisciplinary Research, Chinese Academy of Sciences (CAS), Institute of High Energy Physics, 19B Yuquan Road, Beijing, 100049, China.
authors.forename: Zhifang
authors.initials: Z
authors.lastname: Chai
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Key Lab of Nuclear Radiation and Nuclear Energy Technology, Center for Multidisciplinary Research, Chinese Academy of Sciences (CAS), Institute of High Energy Physics, 19B Yuquan Road, Beijing, 100049, China. huyi@ihep.ac.cn.
authors.forename: Yi
authors.initials: Y
authors.lastname: Hu
mesh_terms: D001360:Avidin / Q000378:metabolism; D057166:High-Throughput Screening Assays; D010748:Phosphopeptides / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D011786:Quality Control; D015203:Reproducibility of Results; D013379:Substrate Specificity; D013601:T-Lymphocytes / Q000201:enzymology
chemical_list: D010748:Phosphopeptides; D001360:Avidin; D017027:Protein Tyrosine Phosphatases
keywords: High-throughput screening; Phosphopeptide microarrays; Protein tyrosine phosphatases; Substrate specificity