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  • Drosophila melanogaster  (2)
  • Key words Metallothionein  (2)
  • Springer  (4)
  • 1
    ISSN: 1573-6857
    Keywords: Drosophila melanogaster ; isofemale lines ; isogroups ; natural populations ; pigmentation ; body size
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Studies of short or medium range geographic variations play an increasing role in ecological genetics, and sensitive techniques are required to detect them. In this respect, two sampling techniques were compared inD. melanogaster. The biological data were provided by the analysis of four natural populations from the same geographic area, Spain (one) and Southern France (three), for four morphometrical traits: abdomen and thoracic pigmentation, and wing and thorax lengths. Traits were measured on wild living females and on their progeny reared in the laboratory at 25°C. For progeny analyses, two techniques were compared: the usual isofemale line technique, sib families issued from a single female, and a new isogroup technique, the progeny produced by a group of 20 wild-collected parents. Large phenotypic variations were observed in wild living flies, corresponding to the unstability of natural environmental conditions during their development. Among laboratory grown flies, variations were much smaller. Between isogroups, differences were small, due to sampling error and some common environment effects. Variations between lines were much greater, thus demonstrating a strong genetic component. When different populations have to be compared, the isogroup technique should be preferred since, for the same amount of work, the lesser variability between groups provides a more precise characterization of the population means.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1327
    Keywords: Key words Metallothionein ; N-Ethylmaleimide ; Alkylation ; Kinetics and mechanism ; 111Cd-NMR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract  The model alkylating agent N-ethylmaleimide (NEM) reacts reversibly at the metal-bound thiolates of Zn7MT and Cd7MT. An unprecedented feature of this reaction is that it approaches equilibrium and requires a large excess of NEM (〉1 mM for 3 μM protein) to drive it to completion. The complex kinetics of the reaction can be followed by monitoring the release of bound metal ions using the metallochromic dyes Zincon (ZI) for Zn7MT and pyridylazoresorcinol for Cd7MT. An initial lag phase is followed by more rapid release of zinc ions. The observed pseudo-first-order rate constants for the two phases are independent of the ZI and Zn7MT concentrations. The complex NEM concentration dependence of each phase, k f, obs=k f 1+k f 2 [NEM] and k s, obs=k s 1+k s 2 [NEM], demonstrates that the forward reactions are second order and the reverse reactions are first order. The alkylation can be reversed using 2-mercaptoethanol to compete for the protein-bound NEM and regenerate the Zn-binding capability of alkylated MT. An explanation of these observations, based on the reversibility of cysteine alkylation by NEM, was developed and tested. The reactions of Cd7MT are less complete than those of Zn7MT and occur more slowly. 111Cd-NMR studies of the partially alkylated 111Cd7MT reveal that reaction with only four equivalents of NEM completely alters the cluster structure and eliminates the spectral signatures of the α and β clusters, although very little cadmium has been removed from the protein. This finding substantiates the proposed kinetic intermediate, a partially alkylated MT with complete or nearly complete retention of the metal ions, and rules out the possibility of cooperative reactions at either cluster.
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  • 3
    ISSN: 1432-1327
    Keywords: Key words Metallothionein ; Peptide-metal complexes ; 111Cd NMR ; Protein folding ; Molecular mechanics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract  The synthetic peptide fragment containing residues 49–61 of rabbit liver metallothionein II (MT-II) (Ac-Ile-Cys-Lys-Gly-Ala-Ser-Asp-Lys-Cys-Ser-Cys-Cys-Ala-COOH), which includes the only sequential four cysteines bound to the same metal ion in Cd7MT, forms a stable, monomeric Cd-peptide complex with 1 : 1 stoichiometry (Cd:peptide) via Cd-thiolate interactions. This represents the first synthesis of a single metal-binding site of MT independent of the domains. The 111Cd NMR chemical shift at 716 ppm indicates that the 111Cd2+ in the metal site is terminally coordinated to four side-chain thiolates of the cysteine residues. The pH of half dissociation for this Cd-peptide derivative, ∼3.3, demonstrates an affinity similar to that for Cd7MT. Molecular mechanics calculations show that the thermodynamically most stable folding for this isolated Cd2+ center has the same counterclockwise chirality (Λ or S) observed in the native holo-protein. These properties are consistent with its proposed role as a nucleation center for cadmium-induced protein folding. However, the kinetic reactivity of the CdS4 structure toward 5,5′-dithiobis(5-nitrobenzoate) (DTNB) and EDTA is greatly increased compared to the complete cluster (α-domain or holo-protein). The rate law for the reaction with DTNB is rate=(k uf +k 1,f +k 2,f [DTNB])[peptide], where k uf=0.15 s–1, k 1,f=2.59×10–3 s–1, and k 2,f=0.88 M–1 s–1. The ultrafast step (uf), observable only by stopped-flow measurement, is unprecedented for mammalian (M7MT) and crustacean (M6MT) holo-proteins or the isolated domains. The accommodation of other metal ions by the peptide indicates a rich coordination chemistry, including stoichiometries of M-peptide for Hg2+, Cd2+, and Zn2+, M2-peptide for Hg2+ and Au+, and (Et3PAu)2-peptide.
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  • 4
    ISSN: 1420-9071
    Keywords: Allozyme polymorphism ; linkage disequilibrium ; wine cellar and field populations ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Over three years, theAdh and α-Gpdh loci have been studied in two cellar populations ofDrosophila melanogaster and in two field populations which were each near to one of the cellars. Analyses of gene frequencies indicate that the divergence among subpopulations is greater in theAdh locus than in the α-Gpdh locus. Selection for or againstAdh S allele acting on theIn(2L)t inversion influences of the α-Gpdh alleles. This phenomenon may contribute to explain the maintenance of theAdh and α-Gpdh polymorphism and of theIn(2L)t inversion.
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