Identification of Two Conserved Residues Involved in Copper Release from Chloroplast PIB-1-ATPases. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Biological Chemistry Année : 2016

Identification of Two Conserved Residues Involved in Copper Release from Chloroplast PIB-1-ATPases.

Résumé

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.
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Dates et versions

hal-01355863 , version 1 (28-05-2020)

Identifiants

Citer

Emeline Sautron, Cécile Giustini, Thuy Van Dang, Lucas Moyet, Daniel Salvi, et al.. Identification of Two Conserved Residues Involved in Copper Release from Chloroplast PIB-1-ATPases.. Journal of Biological Chemistry, 2016, 291 (38), pp.20136-20148. ⟨10.1074/jbc.M115.706978⟩. ⟨hal-01355863⟩
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