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941.
942.
Diego La Mendola Raffaele P. Bonomo Giuseppe Impellizzeri Giuseppe Maccarrone Giuseppe Pappalardo Adriana Pietropaolo Enrico Rizzarelli Valeria Zito 《Journal of biological inorganic chemistry》2005,10(5):463-475
The prion protein (PrPc) is a copper-binding glycoprotein that can misfold into a β-sheet-rich and pathogenic isoform (PrPsc) leading to prion diseases. The first non-mammalian PrPc was identified in chicken and it was found to keep many structural motifs present in mammalian PrPc, despite the low sequence identity (approximately 40%) between the two primary structures. The present paper describes the synthesis and the coordination properties of some hexapeptide fragments (namely, PHNPGY , HNPGYP and NPGYPH) as well as a bishexapeptide (PHNPGYPHNPGY), which encompasses two hexarepeats. The copper(II) complexes were characterized by means of potentiometric, UV–vis, circular dichroism and electron paramagnetic resonance techniques. We also report the synthesis of three hexapeptides (PHNPGF, HNPGFP and NPGFPH), in which one tyrosine was replaced by phenylalanine as well as two bishexapeptides in which either one (PHNPGFPHNPGY and PHNPGYPHNPGF), or two tyrosines were replaced by phenylalanine, in order to check whether tyrosine was involved in copper(II) binding. Overall, the results indicate that the major copper(II) species formed by the chicken PrP dodecapeptides are stabler than the analogous species reported for the peptide fragments containing two octarepeat peptides from the mammalian prion protein. It is concluded that the presence of four prolyl residues, that are break points in copper coordination, induces the metal-assisted formation of macrochelates as well as the formation of binuclear species. Furthermore, it has been shown that the phenolic group is directly involved in the formation of copper binuclear species.Electronic Supplementary Material Supplementary material is available for this article at .This revised version was published online in June 2005 with corrections to the text. The author name LaMendola has been corrected to La Mendola. 相似文献
943.
A seed-feeding weevil, Melanterius ventralis (Coleoptera: Curculionidae), has been introduced into South Africa to supplement a gall wasp,Trichilogaster acaciaelongifoliae (Hymenoptera: Pteromalidae), in the biological control programme against an alien invasive tree, Acacia longifolia (Mimosaceae), from Australia. The gall wasp debilitates most of the flower buds on A. longifolia andreduces seed-set by >95%. The intended rolefor M. ventralis is to destroy theresidual seeds. To achieve this, the gravidfemales need to be able to locate a food sourcethat is both heterogeneously dispersed andfrequently scarce due to damage caused by T.acaciaelongifoliae. Observations showedthat M. ventralis females are meticulousin choosing sites to oviposit so that larvae donot become overcrowded and food limited. Cagetests and field observations revealed thatfemales located pods regardless of density andposition, and that the duration of time spenton branches was proportional to the number ofpods on the branches and to the condition ofpods. The females spent little time onbranches of acacia species other than A.longifolia, but some time was spent onbranches with pods of A. melanoxylon thathad been coated with juice extracted from A.longifolia. All indications are that M.ventralis has the attributes needed to bean excellent supplementary biological controlagent to T. acaciaelongifoliae and theprogress of the weevil continues to be monitored. 相似文献
944.
In the ciliate Euplotes raikovi, two specific isoforms of cAMP-dependent phosphodiesterases were identified, one in the soluble and the other in the particulate fraction of the cell. Their activity was shown to be stimulated by Mg2+, insensitive to Ca2+ and cGMP, and scarcely inhibited by theophylline and 3-isobutyl-1-methyl-xanthine. They appear to be related to some phosphodiesterases of class II of other unicellular organisms in their biochemical features, and their enzymatic activity is up-regulated by elevation of intracellular cAMP level similarly to PDE-4 isoforms of mammals. 相似文献
945.
946.
Fabin H. Acua Lila Ricci Adriana C. Excoffon Mauricio O. Zamponi 《Zoologischer Anzeiger》2004,243(1-2):47-52
Comparative studies on cnidocysts, involving adequate statistical treatment, are very scarce. Classical statistical tests are frequently used assuming normal frequency distributions of capsule lengths, but many distributions are non-normal in acontiarian sea anemones. A traditional choice in these situations are non-parametric tests, although they are not as powerful as parametric tests. An extension of classical methods was developed by some authors; these models, called Generalized Linear Models (GLM), can be used under certain conditions with non-normal data. In view of the properties of our data, that are positive, skewed and with constant coefficient of variation, a GLM with gamma distribution and inverse link function was chosen to analyse the cnidae of acontia from the species Haliplanella lineata, Tricnidactis errans and Anthothoe chilensis. Graphical analysis of residuals showed that these assumptions were reasonable. This method allowed us to avoid transformation of data set and controversial cases in the limit of significance level. For this task, appropriate subroutines in GLIM language were written. In all cases highly significant differences were found between the specimens considered for every species and nematocyst type (b-rhabdoids, p-rhabdoids B1b and p-rhabdoids B2a). 相似文献
947.
Wagner A S Judice Luciano Puzer Simone S Cotrin Adriana K Carmona Graham H Coombs Luiz Juliano Maria A Juliano 《European journal of biochemistry》2004,271(5):1046-1053
The recombinant cysteine peptidases, cruzain from Trypanosoma cruzi and CPB2.8DeltaCTE from Leishmania mexicana, are cathepsin L-like and characteristically endopeptidases. In this study, we characterized the carboxydipeptidase activities of these enzymes and compared them with those of human recombinant cathepsin B and cathepsin L. The analysis used the internally quenched fluorescent peptide Abz-FRFK*-OH and some of its analogues, where Abz is ortho-aminobenzoic acid and K* is (2,4-dinitrophenyl)-epsilon-NH2-lysine. These peptides were demonstrated to be very sensitive substrates, due to the strong quenching effect of K* on the fluorescence of the Abz group. The carboxydipeptidase activity of cruzain was shown to be very similar to that of cathepsin B, while that of CPB2.8DeltaCTE is closer to the carboxydipeptidase activity of cathepsin L. The S2 subsite architecture of cruzain and the nature of the amino acid at the P2 position of the substrates determine its carboxydipeptidase activity and gives further and direct support to the notion that the carboxydipeptidase activity of the papain family cysteine peptidases rely on the S2-P2 interaction [N?gler D. K., Tam, W., Storer, A.C., Krupa, J.C., Mort, J.S. & Menard, R. (1999) Biochemistry38, 4868-4874]. Cruzain and CPB2.8DeltaCTE presented a broad pH-range for both the endo- and exo-peptidase activities, although the later is approximately one order of magnitude lower. This feature, that is not common in related mammalian cysteine peptidases, is consistent with the enzymes being exposed to different environmental conditions and having different locations during parasite development. 相似文献
948.
Adriana A Paulucci Angela M Katsuyama Aurea D Sousa Chuck S Farah 《European journal of biochemistry》2004,271(3):589-600
Tropomyosin is a 284 residue dimeric coiled-coil protein that interacts in a head-to-tail manner to form linear filaments at low ionic strengths. Polymerization is related to tropomyosin's ability to bind actin, and both properties depend on intact N- and C-termini as well as alpha-amino acetylation of the N-terminus of the muscle protein. Nalpha-acetylation can be mimicked by an N-terminal Ala-Ser fusion in recombinant tropomyosin (ASTm) produced in Escherichia coli. Here we show that a recombinant tropomyosin fragment, corresponding to the protein's first 260 residues plus an Ala-Ser fusion [ASTm(1-260)], polymerizes to a much greater extent than the corresponding full-length recombinant protein, despite the absence of the C-terminal 24 amino acids. This polymerization is sensitive to ionic strength and is greatly reduced by the removal of the N-terminal Ala-Ser fusion [nfTm(1-260)]. CD studies show that nonpolymerizable tropomyosin fragments, which terminate at position 260 [Tm(167-260) and Tm(143-260)], as well as Tm(220-284), are able to interact with ASTm(1-142), a nonpolymerizable N-terminal fragment, and that the head-to-tail interactions observed for these fragment pairs are accompanied by a significant degree of folding of the C-terminal tropomyosin fragment. These results suggest that the new C-terminus, created by the deletion, polymerizes in a manner similar to the full-length protein. Head-to-tail binding for fragments terminating at position 260 may be explained by the presence of a greater concentration of negatively charged residues, while, at the same time, maintaining a conserved pattern of charged and hydrophobic residues found in polymerizable tropomyosins from a variety of sources. 相似文献
949.
Recent advances in research on the physiological roles of phosphoinositides in eukaryotic organisms indicate a need to distinguish molecular phosphoinositide species on the basis of their characteristic head groups as well as their glycerolipid moieties. Accurate identification of phosphoinositide species in biological samples poses an analytical challenge, because structurally similar inositol phosphate head groups must be resolved, as must lipid-associated fatty acids. Although intact phosphoinositide species have been successfully analyzed, such analyses employ state-of-the-art liquid chromatography/mass spectrometry and require expensive equipment not accessible to many researchers. Described here is a cost-efficient and reliable alternative developed by adaptation of a combination of classic methods for lipid analysis, thin-layer chromatography and gas chromatography. 相似文献
950.
Sengupta A Carlson BA Hoffmann VJ Gladyshev VN Hatfield DL 《Biochemical and biophysical research communications》2008,365(3):446-452
Selenium is incorporated into proteins as selenocysteine (Sec), which is dependent on its specific tRNA, designated tRNA[Ser]Sec. Targeted removal of the tRNA[Ser]Sec gene (Trsp) in mouse hepatocytes previously demonstrated the importance of selenoproteins in liver function. Herein, analysis of plasma proteins in this Trsp knockout mouse revealed increases in apolipoprotein E (ApoE) that was accompanied by elevated plasma cholesterol levels. The expression of genes involved in cholesterol biosynthesis, metabolism and transport were also altered in knockout mice. Additionally, in two transgenic Trsp mutant mouse lines (wherein only housekeeping selenoprotein synthesis was restored), the expression of ApoE, as well as genes involved in cholesterol biosynthesis, metabolism and transport were similar to those observed in wild type mice. These data correlate with reports that selenium deficiency results in increased levels of ApoE, indicating for the first time that housekeeping selenoproteins have a role in regulating lipoprotein biosynthesis and metabolism. 相似文献