Genome-wide biochemical analysis of Arabidopsis protein phosphatase using a wheat cell-free system.

Plant genome possesses over 100 protein phosphatase (PPase) genes that are key regulators of signal transduction via phosphorylation/dephosphorylation event. Here we report a comprehensive functional analysis of protein serine/threonine, dual-specificity and tyrosine phosphatases using recombinant PPases produced by wheat cell-free protein synthesis system. Eighty-two recombinant PPases were successfully produced using Arabidopsis full-length cDNA as templates. In vitro PPase assay was performed using phosphorylated myelin basic protein as substrate. Among the AtPPases examined, 26 serine/threonine, three dual-specificity and one tyrosine PPases exhibited catalytic activity, including 20 serine/threonine and one dual-specificity PPases that showed in vitro activities for the first time. Our study demonstrates genome-wide biochemical analysis of AtPPases using wheat cell-free system, and provides new information and insights on enzyme activities. STRUCTURED SUMMARY OF PROTEIN INTERACTIONS: PTP1dephosphorylatesMBP by phosphatase assay (View interaction). AtPP2CdephosphorylatesMBP by phosphatase assay (View interaction). POLTEdephosphorylatesMBP by phosphatase assay (View interaction). TOPP8dephosphorylatesMBP by phosphatase assay (View interaction). HAB1dephosphorylatesMBP by phosphatase assay (View interaction). ABI2dephosphorylatesMBP by phosphatase assay (View interaction). At1g34750dephosphorylatesMBP by phosphatase assay (View interaction). At1g43900dephosphorylatesMBP by phosphatase assay (View interaction). At3g15260dephosphorylatesMBP by phosphatase assay (View interaction). At5g53140dephosphorylatesMBP by phosphatase assay (View interaction). At1g18030dephosphorylatesMBP by phosphatase assay (View interaction). At3g06270dephosphorylatesMBP by phosphatase assay (View interaction). At2g25070dephosphorylatesMBP by phosphatase assay (View interaction). At3g02750dephosphorylatesMBP by phosphatase assay (View interaction). At5g10740dephosphorylatesMBP by phosphatase assay (View interaction). at4g26080dephosphorylatesMBP by phosphatase assay (View interaction). At4g28400dephosphorylatesMBP by phosphatase assay (View interaction). At5g06750dephosphorylatesMBP by phosphatase assay (View interaction). At4g31860dephosphorylatesMBP by phosphatase assay (View interaction). At3g17250dephosphorylatesMBP by phosphatase assay (View interaction). At4g38520dephosphorylatesMBP by phosphatase assay (View interaction). At3g05640dephosphorylatesMBP by phosphatase assay (View interaction). At5g66080dephosphorylatesMBP by phosphatase assay (View interaction). At1g79630dephosphorylatesMBP by phosphatase assay (View interaction). At2g30170dephosphorylatesMBP by phosphatase assay (View interaction). At5g24940dephosphorylatesMBP by phosphatase assay (View interaction). Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.