Structural and functional insights of Wilson disease copper-transporting ATPase

Wilson disease is an autosomal recessive disorder of copper metabolism. The gene for this disorder has been cloned and identified to encode a copper-transporting ATPase (ATP7B), a member of a large family of cation transporters, the P-type ATPases. In addition to the core elements common to all P-type ATPases, the Wilson copper-transporting ATPase has a large cytoplasmic N-terminus comprised six heavy metal associated (HMA) domains, each of which contains the copper-binding sequence motif GMT/HCXXC. Extensive studies addressing the functional, regulatory, and structural aspects of heavy metal transport by heavy metal transporters in general, have offered great insights into copper transport by Wilson copper-transporting ATPase. The findings from these studies have been used together with homology modeling of the Wilson disease copper-transporting ATPases based on the X-ray structure of the sarcoplasmic reticulum (SR) calcium-ATPase, to present a hypothetical model of the mechanism of copper transport by copper-transporting ATPases.     

The phylogenetic tree gathering the plant Zn/Cd/Pb/Co P1B-ATPases appears to be structured according to the botanical families

Plant Zn/Cd/Pb/Co P_1B-ATPases (HMAs) play different roles, among which are the control of metal transport from the roots to the shoot and/or from the cytoplasm into the cell vacuole. Transferring the knowledge acquired on HMAs from model species to HMAs from other species requires one to identify orthologues in these other species. Through an extensive screening of the public sequence databases, 96 plant P_1B-ATPases showing orthology to any of the AtHMA1, AtHMA2, AtHMA3 or AtHMA4 isoforms were identified from 32 plant species belonging to 15 botanical families. The number of paralogues within a species varied greatly from species to species, even within a specific botanical family, suggesting that gene duplication events occurred after speciation. The phylogenetic tree gathering the Zn/Cd/Pb/Co P_1B-ATPases was strongly structured according to the botanical family to which the sequences could be related to. In particular, no strict orthology relationship links the Brassicaceae HMAs to the non-Brassicaceae or the Poaceae ones. Recent data showed that the sole rice HMA characterised to date displays different functional properties from the Arabidopsis HMAs. Altogether, data suggest that it might be risky to directly transfer the knowledge acquired through the study of HMAs in model plant species to HMAs from other species.     

Studies on chlorophyll biosynthesis and other things

Our research on chlorophyll biosynthesis, over a period of approximately twenty years, has been described, emphasizing those areas in which our laboratory made significant and timely contributions. References to some of our most important articles are included. Portions of the chlorophyll biosynthetic pathway, in which our own laboratory was not involved, for example, the reduction of protochlorophyllide to chlorophyllide and the phytylation of the latter to yield chlorophyll a, have not been covered in this article. Those events which preceded my involvement with chlorophyll biosynthesis, but which contributed to the formation of my own scientific personality, are mentioned briefly in the Introduction. My non-scientific avocations have been included at the request of the reviewers and Govindjee.     

Metal ion binding to calmodulin: NMR and fluorescence studies

Calmodulin is an important second messenger protein which is involved in a large variety of cellular path-ways.Calmodulin is sensitive to fluctuations in the intracellular Ca levels and is activated by the bindingof four Ca ions. In spite of the important role it plays in signal transduction pathways, it shows a surpris-inglybroad specificity for binding metal ions. Using 15N-Gly biosynthetically-labelled calmodulin, we havestudied the binding of different metal ions to calmodulin, including K+, Na+, Ca, Mg, Zn, Cd, Pb, Hg, Sr, La and Lu, by 1H, 15N HMQC NMR experiments. The effects of these ions on the substrate-bindingability of calmodulin have also been studied by fluorescence spectroscopy of the single tryptophan residue in a 22-residue synthetic peptide encompassing the skeletal muscle myosin light chain kinase calmod-ulin-binding domain. Most of these metal ions can activate a calmodulin target enzyme to some extent,though they bind to calmodulin in a different manner. Mg, which is of direct physiological interest, has adistinct site-preference for calmodulin, as it shows the highest affinity for site I in the N-terminal domain,while the C-terminal sites III and IV are the high affinity binding sites for Ca (as well as for Cd ). At ahigh concentration of Mg and a low concentration of Ca, calmodulin can bind Mg in its N-terminallobe while the C-terminal domain is occupied by Ca; this species could exist in resting cells in which the Mg level significantly exceeds that of Ca. Moreover, our data suggest that the toxicity of Pb-which,like Sr, binds with an equal and high affinity to all four sites-may be related to its capacity to tightlybind and improperly activate calmodulin.     

Decreased abundance and diversity of culturable Pseudomonas spp. populations with increasing copper exposure in the sugar beet rhizosphere

Recent studies have indicated that culturable bacteria constitute highly sensitive bioindicators of metal-induced stress in soil. We report the impact of different copper exposure levels characteristic of contaminated agricultural soils on culturable Pseudomonas spp. in the rhizosphere of sugar beet. We observed that the abundance of Pseudomonas spp. was much more severely affected than that of the general population of culturable heterotrophic bacteria by copper. For diversity assessment, Pseudomonas isolates were divided into operational taxonomic units based on amplified ribosomal DNA restriction analysis and genomic PCR fingerprinting by universally primed PCR. Copper significantly decreased the diversity of Pseudomonas spp. in the rhizosphere and significantly increased the frequency of copper-resistant isolates. Concomitant chemical and biological analysis of copper in the rhizosphere and in bulk soil extracts indicated no rhizosphere effect and a relatively low copper bioavailability in the studied soil, suggesting that the observed effects of copper may occur at lower total concentrations in other soils. We conclude that culturable Pseudomonas sensu stricto constitutes a highly sensitive and relevant bioindicator group for the impact of copper in the rhizosphere habitat, and suggest that continued application of copper to agricultural soils poses a significant risk to successful rhizosphere colonization by Pseudomonas spp.     

Trace Metal Bioaccumulation in Epibenthic Macroinvertebrates (EMI) from the San Pedro Shelf,California

Trace metal/metalloid (As, Ag, Cd, Cr, Cu, Hg, Ni, Pb, Se, Zn) bioaccumulation was measured over a period of ten years (1985–1995) in five species of epibenthic macroinvertebrates (EMI) from the San Pedro Shelf, California. Four of the species are numerical dominants in the Southern California Bight (SCB) and two species are commercially harvested. Among three echinoderms, a predatory sea star yielded highest tissue concentrations compared to a sea cucumber and a sea urchin, and concentrations measured in a mantid shrimp exceeded those from a prawn. Comparison of trace metal bioaccumulation from the study area, including the ocean outfall, and elsewhere indicated that bioaccumulation in local EMI was generally low. Measurements from an outfall station and two reference stations were used to evaluate the spatial and temporal relationship between trace metal bioaccumulation and the wastewater discharge. It was concluded that there was no spatial or temporal relationship between EMI trace metal bioaccumulation and the discharge.     

The Synergistic Effect of Cation and Anion of an Ionic Liquid Additive for Lithium Metal Anodes

Lithium metal anodes are steadily gaining more attention, as their superior specific capacities and low redox voltage can significantly increase the energy density of rechargeable batteries far beyond those of current Li‐ion batteries. Nonetheless, the relevant technology is still in a premature research stage mainly due to the uncontrolled growth of Li dendrites that ceaselessly cause unwanted side reactions with electrolyte. In order to circumvent this shortcoming, herein, an ionic liquid additive, namely, 1‐dodecyl‐1‐methylpyrrolidinium (Pyr1(12)⁺) bis(fluorosulfonyl)imide (FSI^?), for conventional electrolyte solutions is reported. The Pyr1(12)⁺ cation with a long aliphatic chain mitigates dendrite growth via the combined effects of electrostatic shielding and lithiophobicity, whereas the FSI^? anion can induce the formation of rigid solid–electrolyte interphase layers. The synergy between the cation and anion significantly improves cycling performance in asymmetric and symmetric control cells and a full cell paired with an LiFePO₄ cathode. The present study provides a useful insight into the molecular engineering of electrolyte components by manipulating the charge and structures of the involved molecules.     

High Polarity Poly(vinylidene difluoride) Thin Coating for Dendrite‐Free and High‐Performance Lithium Metal Anodes

The high‐polarity β‐phase poly(vinylidene difluoride) (β‐PVDF), which has all trans conformation with F and H atoms located on the opposite sides of the polymer backbone, is demonstrated to be a promising artificial solid‐electrolyte interphase coating on both Cu and Li metal anodes for dendrite‐free Li deposition/stripping and enhanced cycling performance. A thin (≈4 µm) β‐PVDF coating on Cu enables uniform Li deposition/stripping at high current densities up to 5 mA cm^?2, Li‐plating capacity loadings of up to 4 mAh cm^?2, and excellent cycling stability over hundreds of cycles under practical conditions (1 mA cm^?2 with 2 mAh cm^?2). Full cells containing an LiFePO₄ cathode and an anode of either β‐PVDF coated Cu or Li also exhibit excellent cycling stability. The profound effects of the high‐polarity PVDF coating on dendrite suppression are attributed to the electronegative F‐rich interface that favors layer‐by‐layer Li deposition. This study offers a new strategy for the development of dendrite‐free metal anode technology.