Biblioteca Digital

Mechanisms of copper homeostasis are of great interest partly due to their connection to debilitating genetic and neurological disorders. The family of high-affinity copper transporters (Ctr) is responsible for extracellular copper acquisition and internalization in yeast, plants, and mammals, including human. The extracellular domain of the human high-affinity copper transporter (hCtr1) contains essential Cu-binding methionine-rich MXXM and MXM (Mets) motifs that are important for copper acquisition and transport. The hCtr1 extracellular domain also contains potential copper binding histidine (His) clusters, including a high-affinity Cu(II) ATCUN site. As of yet, extracellular His clusters have no established significance for hCtr1 function. We have made model peptides based on the extracellular copper acquisition domain of hCtr1 that is rich in His residues and Mets motifs. The peptides' Cu(I) and Cu(II) binding properties have been characterized by UV-Vis and mass spectrometry. Our findings have been extended to a mouse cell model and we show that His residues are important for hCtr1 function likely because of their contribution to strong copper-binding sites in the hCtr1 extracellular domain responsible for copper acquisition.

Copper's pro-oxidant property is also medicinally promising if it can be harnessed to induce oxidative stress as a cancer chemotherapy strategy. Our lab has designed a photocleavable caged copper complex that can selectively release redox-active copper in response to light. The thermodynamic copper binding properties of these potential chemotherapeutics have been characterized

Copper is an essential trace element required by all aerobic organisms as a co-factor for enzymes involved in normal growth, development and physiology. Ctr1 proteins are members of a highly conserved family of copper importers responsible for copper uptake across the plasma membrane. Mice lacking Ctr1 die during embryogenesis from widespread developmental defects, demonstrating the need for adequate copper acquisition in the development of metazoan organisms via as yet uncharacterized mechanisms. The early lethality of the Ctr1 knockout mouse has made it difficult to study the functions of copper and Ctr1 proteins in metazoan development and physiology. Drosophila melanogaster, a genetically tractable system expresses three Ctr1 genes, Ctr1A, Ctr1B and Ctr1C, and may help to further understand the roles of copper and Ctr1 proteins in metazoan development and physiology. Described here is the characterization of Drosophila Ctr1A.

Localization studies using an affinity purified anti-Ctr1A peptide antibody show Ctr1A is predominantly expressed at the plasma membrane in whole embryos and in larval tissues. Ctr1A is an essential gene in Drosophila as loss-of-function mutants, generated by imprecise p-element excision arrest at early larval stages of development. Inductively coupled plasma mass spectroscopy (ICP-MS) demonstrated that whole body copper levels are reduced in Ctr1A mutants and consequently...

Copper (Cu) is a co-factor that is essential for oxidative phosphorylation, protection from oxidative stress, angiogenesis, signaling, iron acquisition, peptide hormone maturation, and a number of other cellular processes. However, excess copper can lead to membrane damage, protein oxidation, and DNA cleavage. To balance the need for copper with the necessity to prevent accumulation to toxic levels, cells have evolved sophisticated mechanisms to regulate copper acquisition, distribution, and storage. The basic components of these regulatory systems are remarkably conserved in most eukaryotes, and this has allowed the use of a variety of model organisms to further our understanding of how Cu is taken into the cell and utilized.

While the components involved in Cu uptake, distribution, and storage are similar in many eukaryotes, evolution has led to differences in how these processes are regulated. For instance, fungi regulate the components involved in Cu uptake and detoxification primarily at the level of transcription while mammals employ a host of post-translational homeostatic mechanisms. In Saccharomyces cerevisiae, transcriptional responses to copper deficiency are mediated by the copper-responsive transcription factor Mac1. Although Mac1 activates the transcription of genes involved in high affinity copper uptake during periods of deficiency...