title: Biochemical characterization of AtHMA6/PAA1, a chloroplast envelope Cu(I)-ATPase.
abstract: Copper is an essential plant micronutrient playing key roles in cellular processes, among them photosynthesis. In Arabidopsis thaliana, copper delivery to chloroplasts, mainly studied by genetic approaches, is thought to involve two P(IB)-type ATPases: AtHMA1 and AtHMA6/PAA1. The lack of biochemical characterization of AtHMA1 and PAA1, and more generally of plant P(IB)-type ATPases, is due to the difficulty of getting high amounts of these membrane proteins in an active form, either from their native environment or after expression in heterologous systems. In this study, we report the first biochemical characterization of PAA1, a plant copper-transporting ATPase. PAA1 produced in Lactococcus lactis is active, forming an aspartyl phosphate intermediate in the presence of ATP and the adequate metal ion. PAA1 can also be phosphorylated using inorganic phosphate in the absence of transition metal. Both phosphorylation types allowed us to demonstrate that PAA1 is activated by monovalent copper ions (and to a lower extent by silver ions) with an apparent affinity in the micromolar range. In agreement with these biochemical data, we also demonstrate that when expressed in yeast, PAA1 induces increased sensitivities to copper and silver. These data provide the first enzymatic characterization of a P(IB-1)-type plant ATPase and clearly identify PAA1 as a high affinity Cu(I) transporter of the chloroplast envelope.
authors: Laboratoire de Chimie et Biologie des Métaux, UMR5249, CNRS, F-38054 Grenoble, France.
authors: Patrice
authors: P
authors: Catty
authors: Sylvain
authors: S
authors: Boutigny
authors: Roger
authors: R
authors: Miras
authors: Jacques
authors: J
authors: Joyard
authors: Norbert
authors: N
authors: Rolland
authors: Daphné
authors: D
authors: Seigneurin-Berny
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002414:Cations, Monovalent / Q000378:metabolism; D025222:Chloroplast Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002736:Chloroplasts / Q000201:enzymology* / Q000235:genetics; D003300:Copper / Q000378:metabolism*; D017136:Ion Transport / Q000502:physiology; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D002414:Cations, Monovalent; D003300:Copper; D025222:Chloroplast Proton-Translocating ATPases; C477511:PAA1 protein, Arabidopsis