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1.
An investigation of the phosphorescence emission properties of tryptophan (Trp) was carried out in glyceraldehyde-3-phosphate dehydrogenase from yeast and from pig and rabbit muscle. Aided by the external heavy-atom effect of iodide, the dependence on excitation wavelength, and thermal quenching profiles, it was established that the 0,0 vibronic band peaked at 406 nm in the pig and rabbit proteins is made up of overlapping contributions from two Trp residues. In contrast to a previous report [Davis, J.M., & Maki, A.H. (1984) Biochemistry 23, 6249-6256], this implies that even in the muscle enzymes all three aromatic side chains are phosphorescent. Further, when the nature of the local environment of each residue is compared to the crystallographic structure of lobster GPDH, it leads to a complete new assignment of the individual phosphorescence spectra. With each protein, a single Trp, identified as Trp-310, was found to display long-lived phosphorescence at room temperature. The decay of this emission gives evidence of conformational homogeneity among the subunits of the tetrameric molecule. 相似文献
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The decay of Trp phosphorescence of proteins in fluid solutions was shown to provide a sensitive tool for probing the conformational homogeneity of these macromolecules in the millisecond to second time scale. Upon examination of 15 single Trp emitting proteins multiexponential decays were observed in 12 cases, a demonstration that the presence of slowly interconverting conformers in solution is more the norm rather than an exception. The amplitude of preexponential terms, from which the conformer equilibrium is derived, was found to be a sensitive function of solvent composition (buffer, pH, ionic strength and glycerol cosolvent), temperature, and complex formation with substrates and cofactors. In many cases, raising the temperature, a point is reached at which the decay becomes practically monoexponential, meaning that conformer interconversion rates have become commensurate with the triplet lifetime. Estimation of activation free energy barriers to interconversion shows that the large values of DeltaG* are rather similar among polypeptides and that the protein substates involved are sufficiently long-lived to display individual binding/catalytic properties. 相似文献
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Since the discovery that 43 kDa TAR DNA binding protein (TDP-43) is involved in neurodegeneration, studies of this protein have focused on the global effects of TDP-43 expression modulation on cell metabolism and survival. The major difficulty with these global searches, which can yield hundreds to thousands of variations in gene expression level and/or mRNA isoforms, is our limited ability to separate specific TDP-43 effects from secondary dysregulations occurring at the gene expression and various mRNA processing steps. In this review, we focus on two biochemical properties of TDP-43: its ability to bind RNA and its protein-protein interactions. In particular, we overview how these two properties may affect potentially very important processes for the pathology, from the autoregulation of TDP-43 to aggregation in the cytoplasmic/nuclear compartments. 相似文献
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Emanuele Bocchieri 《Plant biosystems》2013,147(5-6):325-336
Abstract The results of two years of collection in a small isle in the southern Sardinia are reported, consisting in a floristic list of 116 entities distributed in 96 genera and 41 families. The biological spectrum of this flora puts in evidence a typical Mediterranean environment, characterized by a marked summer dryness. The ratio: number of entities/surface of the studied site has been compared with that of other small southern Sardinian islands, resulting the highest value. This floristic aspect, as well as differences in the biological spectrum, is interpreted as the result of the presence in the area under study of a desultory link with the mainland, in form of a sandy isthmus. This seems important in breaking the biological balance of the small island. 相似文献
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Valentina Sanghez Maria Teresa Russo Maria Antonietta Ajmone‐Cat Emanuele Cacci Alberto Martire Patrizia Popoli Germana Falcone Flavia Michelini Marco Crescenzi Paolo Degan Luisa Minghetti Margherita Bignami Gemma Calamandrei 《Aging cell》2013,12(4):695-705
The contribution that oxidative damage to DNA and/or RNA makes to the aging process remains undefined. In this study, we used the hMTH1‐Tg mouse model to investigate how oxidative damage to nucleic acids affects aging. hMTH1‐Tg mice express high levels of the hMTH1 hydrolase that degrades 8‐oxodGTP and 8‐oxoGTP and excludes 8‐oxoguanine from both DNA and RNA. Compared to wild‐type animals, hMTH1‐overexpressing mice have significantly lower steady‐state levels of 8‐oxoguanine in both nuclear and mitochondrial DNA of several organs, including the brain. hMTH1 overexpression prevents the age‐dependent accumulation of DNA 8‐oxoguanine that occurs in wild‐type mice. These lower levels of oxidized guanines are associated with increased longevity and hMTH1‐Tg animals live significantly longer than their wild‐type littermates. Neither lipid oxidation nor overall antioxidant status is significantly affected by hMTH1 overexpression. At the cellular level, neurospheres derived from adult hMTH1‐Tg neural progenitor cells display increased proliferative capacity and primary fibroblasts from hMTH1‐Tg embryos do not undergo overt senescence in vitro. The significantly lower levels of oxidized DNA/RNA in transgenic animals are associated with behavioral changes. These mice show reduced anxiety and enhanced investigation of environmental and social cues. Longevity conferred by overexpression of a single nucleotide hydrolase in hMTH1‐Tg animals is an example of lifespan extension associated with healthy aging. It provides a link between aging and oxidative damage to nucleic acids. 相似文献