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W E Rauser

Publications and source records attributed to W E Rauser.

11 recordsLinked to original sources
PubMed1999

X-ray absorption spectroscopy of cadmium phytochelatin and model systems.

Higher plants, algae and some yeasts respond to potentially toxic heavy metals such as cadmium by synthesizing phytochelatins and related cysteine-rich polypeptides. We have used X-ray absorption spectroscopy to study the nature of cadmium binding in such peptides isolated from maize (Zea mays) exposed to low levels of cadmium, and in two synthetic cadmium-peptide complexes, Cd-(gamma-Glu-Cys)3Gly and Cd-(alpha-Glu-Cys)3Gly. We have used the synthetic ions [Cd(SPh)4]2-, [Cd4(SPh)10]2- and [S4Cd10(SPh)16]4-as crystallographically defined models for the cadmium site. The Cd K-edge extended X-ray absorption fine structure (EXAFS) data, together with the Cd K, LI, LII and LIII near-edge spectra, reveal a predominantly tetrahedral coordination of cadmium by sulfur in both the phytochelatin and synthetic peptide complexes. In particular, the Cd LIII-edge lacks a peak at 3534.9 e V which was found to be prominent for oxygen- or nitrogen-coordinated species. The Cd-S distance in the phytochelatin complex is 2.54 A. The Cd K-edge EXAFS does not show any isolated, well-defined Cd-Cd interactions; however, contrary to the conclusion of previous work, their absence is not necessarily indicative of isolated cadmium-thiolate ligation. Evidence from other studies suggests that high static disorder, combined with a large vibrational component, serve to effectively wash out this contribution to the EXAFS. The sulfur K-edge, moreover, shows a low-energy feature both in the phytochelatin and in the synthetic cadmium-peptide complexes which is consistent with sulfide bound in a cluster with cadmium as found for [S4Cd10(SPh)16]4-. This feature strongly suggests the presence of a polynuclear cadmium cluster in maize phytochelatin.

Cadmium↗
PubMed1999

Structure and function of metal chelators produced by plants: the case for organic acids, amino acids, phytin, and metallothioneins.

Plants produce a range of ligands for cadmium (Cd), copper (Cu), nickel (Ni), and zinc (Zn). Cd- and Zn-citrate complexes are prevalent in leaves, even though malate is more abundant. In the xylem sap moving from roots to leaves, citrate and histidine are the principal ligands for Cu, Ni, and Zn. Phosphorus-rich globular bodies in young roots are probably Zn-phytate. Metallothioneins (MTs) are cysteine (Cys)-rich ligands. Plants produce class II MTs (MT-IIs) which differ from the archetypal mammalian MT-I in the location and number of Cys. The Ec protein from wheat embryos has Cys in three domains, binds Zn, and disappears with seedling development. The first 59 amino acids have been sequenced for the protein. Fifty-eight genes for MT-IIs, from a range of plants and tissues, predict proteins with Cys in two domains. Most of the predicted proteins have not been isolated, and their metal binding is poorly documented. Three protein bands, corresponding to six MT genes, have been isolated from Arabidopsis, and the amino acids sequenced for nine fragments. The MT-IIIs are atypical, nontranslationally synthesized polypeptides with variously repeating gamma-glutamylcysteine units. Of the five families known, those with carboxy-terminal glycine are the most widespread among plants, algae, and certain yeasts. A heterogeneous grouping of these molecules form Cd-binding complexes with tetrahedral coordination and a Cd-sulfur interatomic distance of 2.52 A. One complex is cytosolic, the dominant one is vacuolar. Together, they can bind a large proportion of cellular Cd; other ligands may also function. Little is known about the counterpart situation for Cu and Zn.

Cadmium↗
PubMed1995

Three families of thiol peptides are induced by cadmium in maize.

Phytochelatins ((gamma GluCys)nGly) are synthesized from glutathione by plants exposed to metals like Cd2+, Cu2+ and Zn2+. An intracellular complex formed by phytochelatins with Cd2+ and sulfide is thought to detoxify the metal possibly by sequestration in the vacuole. It was found that maize seedlings exposed to Cd2+ produced phytochelatins and two additional families of cysteine-containing peptides, (gamma GluCys)n and (gamma GluCys)nGlu. All thiol peptides with n = 2 and 3 were purified and their structure characterized by tandem mass spectrometry. For maize plants exposed to Cd2+ for 7 days, phytochelatins were synthesized preferentially in the first 24 h whereas the amounts of (gamma GluCys)n and (gamma GluCys)nGlu were the highest thereafter. This was probably due to an initial large pool of glutathione available in control plants compared with a dearth of gamma GluCys and no detectable gamma GluCysGlu. The (gamma GluCys)nGlu peptides were induced exclusively by Cd2+ as they were below the detection limit in control seedlings that contained low amounts of phytochelatins and (gamma GluCys)n. Since the Cys moiety of the peptides is essential for binding Cd2+, a role for accumulated (gamma GluCys)n and (gamma GluCys)nGlu in detoxifying Cd2+ in plants must be considered.

Amino Acid Sequence↗
PubMed1995

Retention of cadmium in roots of maize seedlings. Role of complexation by phytochelatins and related thiol peptides.

Cd from roots of maize was partitioned in seedlings exposed to 3 microM CdSO4 for 1 to 7 d. Most of the root Cd (92-94%) was buffer soluble and provided the classical metal-induced cysteine-rich, high-molecular-weight Cd-binding complex. This complex, however, bound only part of the Cd within the roots, from 19% after 1 d of exposure to 59% by d 7. Three families of peptides formed the Cd-binding complex: (gamma-glutamic acid-cysteine)n-glycine [(gamma-Glu-Cys)n-Gly], or phytochelatins, (gamma-Glu-Cys)n, and (gamma-Glu-Cys)n-Glu. The monothiols gamma-Glu-Cys-Gly (glutathione), gamma-Glu-Cys, and gamma-Glu-Cys-Glu were absent from the complex. The n2 oligomers of any peptide were the least concentrated, whereas the n3 and n4 oligomers increased in the complex with exposure to Cd. By d 7, 75% of (gamma-Glu-Cys)4-Gly, 80% of (gamma-Glu-Cys)4, and 73% of (gamma-Glu-Cys)3-Glu were complexed with Cd. The peptide thiol:Cd molar ratio for the complexes was 1.01 +/- 0.07, as if the minimal amount of thiol was used to bind Cd. Acid-labile sulfide occurred in the complexes from d 1 onward at the low S2-;Cd molar ratio of 0.18 +/- 0.02.

Amino Acid Sequence↗
PubMed1993

gamma-Glutamylcysteinylglutamic acid--a new homologue of glutathione in maize seedlings exposed to cadmium.

Exposure of plants to Cd induces the appearance of several thiols based on glutathione and known as class III metallothioneins (or phytochelatins). A new tripeptide with the structure gamma-GluCysGlu accumulated in roots and shoots of Cd-exposed maize seedlings. This thiol was purified and identified by tandem mass spectrometry. The fragmentation pattern of the maize tripeptide was identical to that of the synthetic compound. Like glutathione, this new tripeptide may serve as a precursor for longer-chain peptides involved in metal detoxification through the formation of Cd-binding complexes.

Amino Acid Sequence↗
PubMed1990

Phytochelatins.

Explore the source record for details and available documents.

Animals↗
PubMed1987

The Cd-binding protein from tomato compared to those of other vascular plants.

Cd-binding protein from tomato roots was partially purified and characterized. The Cd-protein complex was eluted as a single Cd peak from QAE-Sephadex A-25 and purified further on Sephadex G-75 in 1 M KC1 buffer. Circular dichroism measurements showed positive Cotton bands at 232 and 273 nm and a negative band at 253 nm, indicative of Cd-thiolate coordination. The major amino acids were Cys (25.6%), Glx (53.3%), Asx (5.4%) and Gly (12.8%) with no aromatic residues detected. The Cd:Cys ratio was 1:2.4. The material had an apparent molecular mass of 3,000 daltons on gel filtration through Sephadex G-50 fine in 1 M KC1 buffer. The tomato protein resembled the Cd-binding proteins isolated from black bentgrass, maize and cabbage. The presence of Cd-thiolate coordination and the high content of Cys are critical features typical of metallothioneins. The abundance of Glx creates a highly anionic protein which is unlike animal metallothioneins. It is proposed that the Cd-binding proteins from vascular plants be designated phytometallothioneins.

Amino Acids↗
PubMed1968

Ribosomal complexes from an extremely halophilic bacterium and the role of cations.

Concentrated extracts of Halobacterium cutirubrum were prepared at 0 C by gently disrupting cells with a nonionic detergent in a medium containing 3.0 m KCl, 0.5 m NH(4)Cl, and 0.04 m (or more) magnesium acetate and then treating the gelatinous mass with deoxyribonuclease. On KCl-sucrose gradients containing 0.5 m NH(4)Cl and 0.04 m magnesium acetate, these extracts showed 30S and 50S ribosomal subunits plus a flat profile of faster-sedimenting material up to high S values. Only after frozen storage or brief incubation of the extract were 70S ribosomes and distinct classes of small polyribosomes detected. Digestion with ribonuclease converted faster-sedimenting material to 70S particles. The presence of chloramphenicol during preparation of the extracts did not affect these results. The evidence suggests that ribosomal particles exist in these cells as subunits or as polyribosomes but not as 70S ribosomes. To investigate the function of Mg(++) and NH(4) (+) ions in ribosomal complexes from this halophile, concentrated cell extracts and extracts incubated with (14)C-leucine were examined on KCl-sucrose gradients containing different concentrations of these ions. Polyribosomes and the bulk of 70S ribosomes dissociated reversibly to subunits at about 0.01 m Mg(++), whereas a small fraction of the 70S particles, including those which in vitro incorporated (14)C-leucine into nascent protein, dissociated only below 1 mm Mg(++). Below this concentration of Mg(++), nascent protein remained attached to the 50S subunit even at 0.04 mm Mg(++) in the presence of 0.35 to 0.5 m NH(4)Cl. Nascent protein, presumably as peptidyl-transfer ribonucleic acid, dissociated reversibly from 50S subunits only at 0.04 mm Mg(++) and 0.1 m or less NH(4) (+). Thus, the stability of polyribosomes from H. cutirubrum depends specifically on both Mg(++) and NH(4) (+) ions.

Ammonium Chloride↗