title: Identification of Two Conserved Residues Involved in Copper Release from Chloroplast PIB-1-ATPases.
abstract: Copper is an essential transition metal for living organisms. In the plant model Arabidopsis thaliana, half of the copper content is localized in the chloroplast, and as a cofactor of plastocyanin, copper is essential for photosynthesis. Within the chloroplast, copper delivery to plastocyanin involves two transporters of the PIB-1-ATPases subfamily: HMA6 at the chloroplast envelope and HMA8 in the thylakoid membranes. Both proteins are high affinity copper transporters but share distinct enzymatic properties. In the present work, the comparison of 140 sequences of PIB-1-ATPases revealed a conserved region unusually rich in histidine and cysteine residues in the TMA-L1 region of eukaryotic chloroplast copper ATPases. To evaluate the role of these residues, we mutated them in HMA6 and HMA8. Mutants of interest were selected from phenotypic tests in yeast and produced in Lactococcus lactis for further biochemical characterizations using phosphorylation assays from ATP and Pi Combining functional and structural data, we highlight the importance of the cysteine and the first histidine of the CX3HX2H motif in the process of copper release from HMA6 and HMA8 and propose a copper pathway through the membrane domain of these transporters. Finally, our work suggests a more general role of the histidine residue in the transport of copper by PIB-1-ATPases.
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and.
authors: Emeline
authors: E
authors: Sautron
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and.
authors: Cécile
authors: C
authors: Giustini
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and.
authors: ThuyVan
authors: T
authors: Dang
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and.
authors: Lucas
authors: L
authors: Moyet
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and.
authors: Daniel
authors: D
authors: Salvi
authors: the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the CNRS, Laboratoire de Chimie et Biologie des Métaux, UMR 5249, F-38054 Grenoble, France.
authors: Serge
authors: S
authors: Crouzy
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and.
authors: Norbert
authors: N
authors: Rolland
authors: the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the CNRS, Laboratoire de Chimie et Biologie des Métaux, UMR 5249, F-38054 Grenoble, France patrice.catty@cea.fr.
authors: Patrice
authors: P
authors: Catty
authors: From the CNRS, Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, F-38054 Grenoble, France, the Université Grenoble Alpes, F-38054 Grenoble, France, the CEA, DSV, BIG, F-38054 Grenoble, France, the INRA, LPCV, UMR 1417, F-38054 Grenoble, France, and daphne.berny@univ-grenoble-alpes.fr.
authors: Daphné
authors: D
authors: Seigneurin-Berny
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003300:Copper / Q000737:chemistry* / Q000378:metabolism; D006639:Histidine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D060431:Thylakoid Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020524:Thylakoids / Q000201:enzymology* / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; C000611728:HMA8 protein, Arabidopsis; D011994:Recombinant Proteins; D060431:Thylakoid Membrane Proteins; D006639:Histidine; D003300:Copper; D000251:Adenosine Triphosphatases; C000611727:HMA6 protein, Arabidopsis
keywords: ATPase; Arabidopsis; PIB-1-ATPases; chloroplast; copper release site; copper transport; plant biochemistry