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Isolation and characterization of a self-sufficient one-domain protein. (Cd)-metallothionein from Eisenia foetida. 总被引:4,自引:0,他引:4
C Gruber S Stürzenbaum P Gehrig R Sack P Hunziker B Berger R Dallinger 《European journal of biochemistry》2000,267(2):573-582
Earthworms have been shown to accumulate trace elements in general, and particularly high amounts of metal ions such as cadmium, copper and zinc. The earthworm's response to metal contamination has been linked to the induction and expression of metallothionein (MT) proteins, a detoxification strategy analogous to that found in other biological systems. The present study focuses on an inducible Cd-MT isolated from the compost-dwelling brandling worm Eisenia foetida (Savigny). A full characterization of the protein (including protein induction, MT cDNA, amino-acid sequence and metal stoichiometry) revealed a new dimension of knowledge to the molecular genetic information available to date. Whereas the elucidated cDNA codes for a putative protein which possesses 80 amino-acid residues, the characterized protein bears only 41 amino acids. The isolated product has evidently attained its size and shape by cleavage near the N-terminal site and at the linker region between the two putative metal-binding domains of the translated product, yielding a small MT moiety which contains 12 Cys residues (including one triple Cys-motif) binding four cadmium ions. It can be shown that the isolated MT molecule represents a self-sufficient one-domain MT which is stable in vitro. The isolation of the single-domain MT peptide raises the question about the method of formation and significance in vivo of such small MT moieties from tissues of E. foetida and possibly other terrestrial invertebrates. In this respect, two hypotheses are discussed: firstly, the possibility of formation of small MT peptides due to enzymatic cleavage of the intact protein during the process of preparation and isolation; and secondly, the possibility of deliberate post-translational processing of the translated gene product to yield functional one-domain MT moieties. 相似文献
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Metallothioneins in terrestrial invertebrates: structural aspects, biological significance and implications for their use as biomarkers. 总被引:5,自引:0,他引:5
R Dallinger B Berger C Gruber P Hunziker S Stürzenbaum 《Cellular and molecular biology, including cyto-enzymology》2000,46(2):331-346
During the last few years the subject of metallothioneins (MTs) in terrestrial invertebrates has gained increasing attention. One reason for this may be that terrestrial invertebrates provide new insights into the biological diversity of MTs, with the potential of discovering alternative models of structural and functional relationships. Four groups of terrestrial invertebrates have been studied in detail, namely nematodes, insects, snails and earthworms, with the present article focusing on MTs from the latter two groups. Snails are interesting because they possess distinct MT isoforms involved in different metal-specific tasks. In the Roman snail (Helix pomatia), for example, one isoform is predominantly expressed in the midgut gland, accounting for the accumulation, binding and detoxification of cadmium. The second isoform, which is present in the snail's mantle, is substantially different regarding its primary structure. Furthermore, it binds nearly exclusively copper, and thus is probably involved in the homeostatic regulation of essential trace elements. Earthworm MTs merit our attention because of another peculiarity: they seem to be much more unstable than snail MTs, particularly under conventional conditions of preparation. The cDNA of the brandling worm (Eisenia foetida), for instance, codes for a putative MT, which is about twice the size of the actual protein. The isolated MT peptide binds four Cd2+ ions and represents a one-domain MT entity that is stable and functional in vitro. This strongly suggests that earthworm MTs are either posttranslationally modified, or subjected to enzymatic cleavage during preparation. Both snail and earthworm MTs are inducible by metal exposure, especially by cadmium, thus supporting the idea of using them as potential biomarkers for environmental metal pollution. Whilst snail MTs have already been tested in this respect with some success, the use of earthworm MTs as biomarkers still remains to be evaluated, especially in the light of the unknown significance of their posttranslational instability. 相似文献
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C. elegans metallothioneins: new insights into the phenotypic effects of cadmium toxicosis 总被引:11,自引:0,他引:11
Swain SC Keusekotten K Baumeister R Stürzenbaum SR 《Journal of molecular biology》2004,341(4):951-959
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S R Stürzenbaum 《BioTechniques》1999,27(1):50-52
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Earthworm pre-procarboxypeptidase: a copper responsive enzyme 总被引:2,自引:0,他引:2
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Spanier B Stürzenbaum SR Holden-Dye LM Baumeister R 《Journal of molecular biology》2005,352(2):429-437
The control of signal peptide activity by cell surface proteases is one of the main factors that regulate the development and behaviour of organisms. In mammals, neprilysins (NEPs) are known to play a key role in these processes and their inactivation can initiate cellular disorganisation, which in turn may lead to prostate cancer or Hirschsprung disease. Although the proteome of the nematode Caenorhabditis elegans has been intensively studied, very little is known about the function of neprilysins. ZK20.6 (NEP-1), the C.elegans protein with highest identity to mammalian neprilysins, is a 753 amino acid residue protein that displays all neprilysin-typical characteristics, including a short intracellular domain, a transmembrane domain and a long extracellular active domain. Here we show that the expression pattern of nep-1 is limited to pharyngeal cells and a single head neuron. Compared to wild-type, the locomotion of nep-1 knockout animals is significantly impaired, a phenotype that can be rescued by the extrachromosomal re-introduction of nep-1. This suggests that this enzyme plays an important role in the regulation of nematode locomotion. Finally, electrophysiological recording of the pharyngeal activity showed a high sensitivity of the nep-1 pharynx to serotonin (5-HT) and to the neuropeptide AF1 (C.elegans FLP-8), indicating that NEP-1 is a central component that controls the neuronal innervation of pharyngeal pumping in C.elegans. 相似文献