Cellular Distribution Of Metals Research Articles

The role of post-transcriptional modulators of metalloproteins in response to metal deficiencies.

Copper and iron proteins have a wide range of functions in living organisms. Metal assembly into metalloproteins is a complex process, where mismetalation is detrimental and energy consuming to cells. Under metal deficiency, metal distribution is expected to reach a metalation ranking, prioritizing essential versus dispensable metalloproteins, while avoiding interference with other metals and protecting metal-sensitive processes. In this review, we propose that post-transcriptional modulators of metalloprotein mRNA (ModMeR) are good candidates in metal prioritization under metal-limited conditions. ModMeR target high quota or redundant metalloproteins and, by adjusting their synthesis, ModMeR act as internal metal distribution valves. Inappropriate metalation of ModMeR targets could compete with metal delivery to essential metalloproteins and interfere with metal-sensitive processes, such as chloroplastic photosynthesis and mitochondrial respiration. Regulation of ModMeR targets could increase or decrease the metal flow through interconnected pathways in cellular metal distribution, helping to achieve adequate differential metal requirements. Here, we describe and compare ModMeR that function in response to copper and iron deficiencies. Specifically, we describe copper-miRNAs from Arabidopsis thaliana and diverse iron ModMeR from yeast, mammals, and bacteria under copper and iron deficiencies, as well as the influence of oxidative stress. Putative functions derived from their role as ModMeR are also discussed.

The effect of metalloprotein inhibitors on cellular metal ion content and distribution.

With metalloproteins garnering increased interest as therapeutic targets, designing target-specific metalloprotein inhibitors (MPi) is of substantial importance. However, in many respects, the development and evaluation of MPi lags behind that of conventional small molecule therapeutics. Core concerns around MPi, such as target selectivity and potential disruption of metal ion homeostasis linger. Herein, we used a suite of analytical methods, including energy-dispersive X-ray spectroscopy (EDX), inductively coupled plasma atomic emission spectroscopy (ICP-OES), and synchrotron X-ray fluorescence microscopy (SXRF) to investigate the effect of several MPi on cellular metal ion distribution and homeostasis. The results reveal that at therapeutically relevant concentrations, the tested MPi have no significant effects on cellular metal ion content or distribution. In addition, the affinity of the metal-binding pharmacophore (MBP) utilized by the MPi does not have a substantial influence on the effect of the MPi on cellular metal distribution. These studies provide an important, original data set indicating that metal ion homeostasis is not notably perturbed by MPi, which should encourage the development of and aid in designing new MPi, guide MBP selection, and clarify the effect of MPi on the 'metallome'.

Stimulus-Responsive Prochelators for Manipulating Cellular Metals.

Metal ions are essential for a wide range of physiological processes, but they can also be toxic if not appropriately regulated by a complex network of metal trafficking proteins. Intervention in cellular metal distribution with small-molecule or peptide chelating agents has promising therapeutic potential to harness metals to fight disease. Molecular outcomes associated with forming metal-chelate interactions in situ include altering the concentration and subcellular metal distribution, inhibiting metalloenzymes, enhancing the reactivity of a metal species to elicit a favorable biological response, or passivating the reactivity of a metal species to prevent deleterious reactivity. The systemic administration of metal chelating agents, however, raises safety concerns due to the potential risks of indiscriminate extraction of metals from critical metalloproteins and inhibition of metalloenzymes. One can estimate that chelators capable of complexing metal ions with dissociation constants in the submicromolar range are thermodynamically capable of extracting metal ions from some metalloproteins and disrupting regular function. Such dissociation constants are easily attainable for multidentate chelators interacting with first-row d-block metal cations in relevant +1, + 2, and +3 oxidation states. To overcome this challenge of indiscriminate metal chelation, we have pursued a prodrug strategy for chelating agents in which the resulting "prochelator" has negligible metal binding affinity until a specific stimulus generates a favorable metal binding site. The prochelator strategy enables conditional metal chelation to occur preferentially in locations affected by disease- or therapy-associated stimuli, thereby minimizing off-target metal chelation. Our design of responsive prochelators encompasses three general approaches of activation: the "removal" approach operates by eliminating a masking group that blocks a potential metal chelation site to reveal the complete binding site under the desired conditions; the molecular "switch" approach involves a reversible conformational change between inactive and active forms of a chelator with differential metal binding affinity under specific conditions; and the "addition" approach adds a new ligand donor arm to the prochelator to constitute a complete metal chelation site. Adopting these approaches, we have created four categories of triggerable prochelators that respond to (1) reactive oxygen species, (2) light, (3) specific enzymes, and (4) biological regulatory events. This Account highlights progress from our group on building prochelators that showcase these four categories of responsive metal chelating agents for manipulating cellular metals. The creation and chemical understanding of such stimulus-responsive prochelators enables exciting applications for understanding the cell biology of metals and for developing therapies based on metal-dependent processes in a variety of diseases.

Cellular partitioning of nanoparticulate versus dissolved metals in marine phytoplankton.

Discharges of metal oxide nanoparticles into aquatic environments are increasing with their use in society, thereby increasing exposure risk for aquatic organisms. Separating the impacts of nanoparticle from dissolved metal pollution is critical for assessing the environmental risks of the rapidly growing nanomaterial industry, especially in terms of ecosystem effects. Metal oxides negatively affect several species of marine phytoplankton, which are responsible for most marine primary production. Whether such toxicity is generally due to nanoparticles or exposure to dissolved metals liberated from particles is uncertain. The type and severity of toxicity depends in part on whether phytoplankton cells take up and accumulate primarily nanoparticles or dissolved metal ions. We compared the responses of the marine diatom, Thalassiosira weissflogii, exposed to ZnO, AgO, and CuO nanoparticles with the responses of T. weissflogii cells exposed to the dissolved metals ZnCl2, AgNO3, and CuCl2 for 7 d. Cellular metal accumulation, metal distribution, and algal population growth were measured to elucidate differences in exposure to the different forms of metal. Concentration-dependent metal accumulation and reduced population growth were observed in T. weissflogii exposed to nanometal oxides, as well as dissolved metals. Significant effects on population growth were observed at the lowest concentrations tested for all metals, with similar toxicity for both dissolved and nanoparticulate metals. Cellular metal distribution, however, markedly differed between T. weissflogii exposed to nanometal oxides versus those exposed to dissolved metals. Metal concentrations were highest in the algal cell wall when cells were exposed to metal oxide nanoparticles, whereas algae exposed to dissolved metals had higher proportions of metal in the organelle and endoplasmic reticulum fractions. These results have implications for marine plankton communities as well as higher trophic levels, since metal may be transferred from phytoplankton through food webs vis à vis grazing by zooplankton or other pathways.

Imaging of metals in biological tissue by laser ablation inductively coupled plasma mass spectrometry (LA–ICP–MS): state of the art and future developments

Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is well established as a sensitive trace and ultratrace analytical technique with multielement capability for bioimaging of metals and studying metallomics in biological and medical tissue. Metals and metalloproteins play a key role in the metabolism and formation of metal-containing deposits in the brain but also in the liver. In various diseases, analysis of metals and metalloproteins is essential for understanding the underlying cellular processes. LA-ICP-MS imaging (LA-ICP-MSI) combined with other complementary imaging techniques is a sophisticated tool for investigating the regional and cellular distribution of metals and related metal-containing biomolecules. On the basis of successful routine techniques for the elemental bioimaging of cryosections by LA-ICP-MSI with a spatial resolution between 200 and ~10 µm, the further development used online laser microdissection ICP-MSI to study the metal distribution in small biological sample sections (at the cellular level from 10 µm to the submicrometer range). The use of mass spectrometric imaging of metals and also nonmetals is demonstrated on a series of biological specimens. This article discusses the state of the art of bioimaging of metals in thin biological tissue sections by LA-ICP-MSI with spatial resolution at the micrometer scale, future developments and prospects for quantitative imaging techniques of metals in the nanometer range. In addition, combining quantitative elemental imaging by LA/laser microdissection-ICP-MSI with biomolecular imaging by matrix-assisted laser desorption/ionization-MSI will be challenging for future life science research.

Cellular Distribution of Metals in a Liverwort and a Moss Transplanted to Two Streams of Differing Acidity

To investigate the consequences of acidification and metal accumulation on the biology of aquatic bryophytes, the acid-tolerant liverwort Scapania undulata (L.) Dum. and the acid-sensitive moss Rhynchostegium riparioides (Hedw.) Cardot were transplanted from one stream to two other streams of differing acidity (pH 5.20 and 6.38). The bryophytes were collected in a circumneutral (pH 6.57) stream in the Vosges Mountains. Metal accumulation was semi-quantitatively measured in shoots by energy dispersive TEM X-ray spectroscopy (EDXS). After 1 month, the two species remained green without alteration signs. Although no marked ultrastructural damage was observed in either species, some cells seemed to be necrotic, with flattened chloroplasts, in R. riparioides. Lipid droplet accumulation was observed in some leaf cells of S.undulata when transplanted to the most acidic stream. Metal was mostly localised in the cell wall, and was only sometimes detected in small vacuoles. Under acidic conditions, R. riparioides showed the highest relative amount of Al and the lowest amount of Fe, whereas the acid-tolerant bryophyte species S. undulata contained more Fe and less Al. The capability to limit the uptake of metals into the cytoplasm varies according to the bryophyte species. This could be an explanation of the tolerance of S. undulata to acidification.

A biomonitoring method using the cellular distribution of metals in moss

Samples of the feather moss Hylocomium splendens, were subjected to a sequential elution procedure in order to quantify the metal content of the intercellular, extracellular exchangeable, intracellular and particulate fractions. Measurements were made on segments of different age and with increasing distance from a heavy metal pollution source. The effect of desiccation and storage time following field sampling was shown to modify the cellular location of elements, but rehydration could partly reverse such responses. Ni and Pb were shown to be equally effective for the elution of extracellular exchangeable Zn supplied under laboratory conditions. The value of the sequential elution procedure for heavy metal biomonitoring studies is discussed.

Cellular metal distribution in the Pacific oyster, Crassostrea gigas (Thun.) determined by quantitative X-ray microprobe analysis

Tissues of Pacific oysters, Crassostrea gigas (Thun.), from the metal-rich Derwent Estuary, Tasmania, were examined using electron microscopy and the cellular metal concentrations measured by X-ray microanalysis. These data showed that 1. (1) all tissues contained elevated copper and zinc concentrations when compared with other bivalves (e.g. Mytilus), 2. (2) with the exception of reproductive tissues, calcium, iron, copper and zinc were present in the tertiary lysosomes, 3. (3) copper and zinc were selectively accumulated in membrane-limited vesicles of blood amoebocytes to cellular concentrations of ≈ 70 mM Cu and 390 mM Zn and 4. (4) these amoebocytes contain > 90% of the body copper and zinc.