title: Expression of the Arabidopsis thaliana AtJ2 cochaperone protein in Pichia pastoris.
abstract: A vector was constructed for intracellular expression of the Arabidopsis thaliana DnaJ homologue AtJ2 in the methylotrophic yeast Pichia pastoris. The vector includes DNA encoding an amino-terminal histidine-tag, to simplify protein purification. Shake-flask cultures could be induced to produce approximately 250 mg/ L of AtJ2. Purified recombinant AtJ2 was able to stimulate the ATPase activities of both the Escherichia coli and Zea mays cytoplasmic Stress70 chaperone proteins five- to ninefold. The carboxy terminus of AtJ2 is -CAQQ, a protein farnesylation motif. When transformed P. pastoris was induced to synthesize AtJ2 in the presence of [(3)H]mevalonolactone, radioactivity was incorporated into the protein, suggesting farnesylation.
authors.affiliation: Institute of Agro-Physics, Plant Physiology and Biochemistry, Hebei Academy of Agricultural Sciences, Shijiazhuang, People's Republic of China.
authors.forename: R
authors.initials: R
authors.lastname: Zhou
authors.forename: B
authors.initials: B
authors.lastname: Kroczyńska
authors.forename: J A
authors.initials: JA
authors.lastname: Miernyk
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry; D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins; D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006639:Histidine / Q000737:chemistry; D018832:Molecular Chaperones / Q000378:metabolism*; D010843:Pichia / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: C494633:ATJ2 protein, Arabidopsis; D029681:Arabidopsis Proteins; C494635:DnaJ protein, E coli; D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D006639:Histidine; D000251:Adenosine Triphosphatases