title: Genome-wide biochemical analysis of Arabidopsis protein phosphatase using a wheat cell-free system.
abstract: 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.
authors.affiliation: Cell-Free Science and Technology Research Center, Ehime University, Matsuyama, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Takahashi
authors.forename: Akihiko
authors.initials: A
authors.lastname: Ozawa
authors.forename: Keiichirou
authors.initials: K
authors.lastname: Nemoto
authors.forename: Akira
authors.initials: A
authors.lastname: Nozawa
authors.forename: Motoaki
authors.initials: M
authors.lastname: Seki
authors.forename: Kazuo
authors.initials: K
authors.lastname: Shinozaki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takeda
authors.forename: Yaeta
authors.initials: Y
authors.lastname: Endo
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Sawasaki
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D002474:Cell-Free System; D018744:DNA, Plant / Q000235:genetics; D018745:Genome, Plant; D010749:Phosphoprotein Phosphatases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D014908:Triticum / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D018744:DNA, Plant; D011994:Recombinant Proteins; D010749:Phosphoprotein Phosphatases