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

Sautron, Emeline; Giustini, Cécile; Dang, Thuy Van; Moyet, Lucas; Salvi, Daniel; Crouzy, Serge; Rolland, Norbert; Catty, Patrice; Seigneurin-Berny, Daphné

doi:10.1074/jbc.m115.706978

Cited by 8 publications

(3 citation statements)

References 40 publications

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“…Their presence in high concentrations, however, tends to be detrimental to cells ( 33 ). The Cu-histidine interaction highlighted by our model has been well studied across many forms of life ( 34 – 40 ). Copper is known to play diverse structural and catalytic functions owing to its ability to exist either in a reduced (Cu + ) state with affinity for thiol and thioether groups or in an oxidized (Cu 2+ ) state with more likely coordination for the oxygen or imidazole nitrogen group of amino acids, including histidine ( 41 ).…”

Section: Discussionmentioning

confidence: 98%

Applying Statistical Design of Experiments To Understanding the Effect of Growth Medium Components on Cupriavidus necator H16 Growth

Azubuike

Edwards

Gatehouse

et al. 2020

Appl Environ Microbiol

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Cupriavidus necator H16 is gaining significant attention as a microbial chassis for range of biotechnological applications. Whilst the bacterium is a major producer of bioplastics, its lithoautotrophic and versatile metabolic capabilities make the bacterium a promising microbial chassis for biofuels and chemicals using renewable resources. It remains necessary to develop appropriate experimental resources to permit controlled bioengineering and system optimization of this microbe. Here we employed statistical design of experiments to gain understanding of the impact of components of defined media on C. necator growth and built a model that can predict the bacterium's cell density based on media components. This highlighted media components, and interaction between components, having the most effect on growth: fructose, amino acids, trace elements, CaCl2 and Na2HPO4 contributed significantly to growth (t < -1.65 or > 1.65); copper and histidine were found to interact and must be balanced for robust growth. Our model was experimentally validated and found to correlate well, r2 = 0.85. Model validation at large culture scales showed correlations between our model predicted growth ranks and experimentally determined ranks at 100 mL in shake flasks (ρ = 0.87) and 1 L in bioreactor (ρ = 0.90). Our approach provides valuable and quantifiable insights on the impact of media components on cell growth and can be applied to model other C. necator responses that are crucial for its deployment as a microbial chassis. This approach can be extended to other non-model microbes of medical and industrial biotechnological importance. Importance Chemically defined media (CDM) for cultivation of C. necator vary in components and compositions. This lack of consensus makes it difficult optimizing new processes for the bacterium. This study employed statistical design of experiments (DOE) to understand how basic components of defined media affect C. necator growth. Our growth model predicts that C. necator can be cultivated to high-cell-density with components held at low concentrations, arguing that CDM for large scale cultivation of the bacterium for industrial purposes will be economically competitive. Although existing CDM for the bacterium are without amino acids, addition of few amino acids to growth medium shortened lag phase of growth. The interactions highlighted by our growth model shows how factors can interact with each other during a process to positively or negatively affect process output. This approach is efficient, relying on few well-structured experimental runs to gain maximum information on a biological process, growth.

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Section: Discussionmentioning

confidence: 98%

Applying Statistical Design of Experiments To Understanding the Effect of Growth Medium Components on Cupriavidus necator H16 Growth

Azubuike

Edwards

Gatehouse

et al. 2020

Appl Environ Microbiol

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“…CpeHMA6 showed a high upregulation in leaf tissue under Cu treatment and in root and flower tissues under As stress. It was observed from the phylogenetic tree that CpHMA6 is orthologous to AtHMA8 from Arabidopsis, which is related to the Cu transport through the thylakoid membrane [69][70][71]. Cu is an essential metal due to its function as an enzyme cofactor for a number of physiological processes [72].…”

Section: Discussionmentioning

confidence: 99%

A Genome-Wide Identification and Comparative Analysis of the Heavy-Metal-Associated Gene Family in Cucurbitaceae Species and Their Role in Cucurbita pepo under Arsenic Stress

Flores-Iga,

Lopez-Ortiz,

Gracia-Rodriguez

et al. 2023

Genes

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The heavy-metal-associated (HMA) proteins are a class of PB1-type ATPases related to the intracellular transport and detoxification of metals. However, due to a lack of information regarding the HMA gene family in the Cucurbitaceae family, a comprehensive genome-wide analysis of the HMA family was performed in ten Cucurbitaceae species: Citrullus amarus, Citrullus colocynthis, Citrullus lanatus, Citrullus mucosospermus, Cucumis melo, Cucumis sativus, Cucurbita maxima, Cucurbita moschata, Cucurbita pepo, and Legenaria siceraria. We identified 103 Cucurbit HMA proteins with various members, ranging from 8 (Legenaria siceraria) to 14 (Cucurbita pepo) across species. The phylogenetic and structural analysis confirmed that the Cucurbitaceae HMA protein family could be further classified into two major clades: Zn/Co/Cd/Pb and Cu/Ag. The GO-annotation-based subcellular localization analysis predicted that all HMA gene family members were localized on membranes. Moreover, the analysis of conserved motifs and gene structure (intron/exon) revealed the functional divergence between clades. The interspecies microsynteny analysis demonstrated that maximum orthologous genes were found between species of the Citrullus genera. Finally, nine candidate HMA genes were selected, and their expression analysis was carried out via qRT-PCR in root, leaf, flower, and fruit tissues of C. pepo under arsenic stress. The expression pattern of the CpeHMA genes showed a distinct pattern of expression in root and shoot tissues, with a remarkable expression of CpeHMA6 and CpeHMA3 genes from the Cu/Ag clade. Overall, this study provides insights into the functional analysis of the HMA gene family in Cucurbitaceae species and lays down the basic knowledge to explore the role and mechanism of the HMA gene family to cope with arsenic stress conditions.

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“…One may speculate that target proteins to be loaded with copper ions in the Golgi obtain Cu(I) ions from these exit sites via direct protein-protein interactions, although no such studies have yet been performed. Notably, in corresponding chloroplast P 1B-1 -type ATPases HMA6 and HMA8, conserved Cu(I)-interacting (here, His and Cys residues) positions have also been described in transmembrane helix 1 and the following lumen-protruding peptide segment that appear important for Cu(I) release in the chloroplasts (55).…”

Section: Discussionmentioning

confidence: 99%

A Luminal Loop of Wilson Disease Protein Binds Copper and Is Required for Protein Activity

Köhn

Shanmugavel

et al. 2018

Biophysical Journal

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The copper-transporting ATPase ATP7B is essential for loading of copper ions to copper-dependent enzymes in the secretory pathway; its inactivation results in Wilson disease. In contrast to copper-ion uptake by the cytoplasmic domains, ATP7B-mediated copper-ion release in the Golgi has not been explored yet. We demonstrate here that a luminal loop in ATP7B, rich in histidine/methionine residues, binds reduced copper (Cu(I)) ions, and identified copper-binding residues play an essential role in ATP7B-mediated metal ion release. NMR experiments on short-peptide models demonstrate that three methionine and two histidine residues are specifically involved in Cu(I) ion binding; with these residues replaced by alanines, no Cu(I) ion interaction is detected. Although more than one Cu(I) ion can interact with the wild-type peptide, removing either all histidine or all methionine residues reduces the stoichiometry to one Cu(I) ion binding per peptide. Using a yeast complementation assay, we show that for efficient copper transport by full-length ATP7B, the complete set of histidine and methionine residues in the lumen loop are required. The replacement of histidine/methionine residues by alanines does not perturb overall ATP7B structure, as the localization of ATP7B variants in yeast cells matches that of the wild-type protein. Thus, in similarity to ATP7A, ATP7B also appears to have a luminal "exit" copper ion site.

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

Cited by 8 publications

References 40 publications

Applying Statistical Design of Experiments To Understanding the Effect of Growth Medium Components on Cupriavidus necator H16 Growth

Applying Statistical Design of Experiments To Understanding the Effect of Growth Medium Components on Cupriavidus necator H16 Growth

A Genome-Wide Identification and Comparative Analysis of the Heavy-Metal-Associated Gene Family in Cucurbitaceae Species and Their Role in Cucurbita pepo under Arsenic Stress

A Luminal Loop of Wilson Disease Protein Binds Copper and Is Required for Protein Activity

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