TY - CHAP
T1 - High-throughput screening of substrate specificity for protein tyrosine phosphatases (PTPs) on phosphopeptide microarrays
AU - Gao, Liqian
AU - Lee, Su Seong
AU - Chen, Jun
AU - Sun, Hongyan
AU - Zhao, Yuliang
AU - Chai, Zhifang
AU - Hu, Yi
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2016.
PY - 2016/5/1
Y1 - 2016/5/1
N2 - 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.
AB - 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.
KW - High-throughput screening
KW - Phosphopeptide microarrays
KW - Protein tyrosine phosphatases
KW - Substrate specificity
UR - http://www.scopus.com/inward/record.url?scp=84965053644&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84965053644&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-3136-1_13
DO - 10.1007/978-1-4939-3136-1_13
M3 - Chapter
C2 - 26614076
AN - SCOPUS:84965053644
T3 - Methods in Molecular Biology
SP - 181
EP - 196
BT - Methods in Molecular Biology
PB - Humana Press
ER -