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51.
Identification of a cysteine residue at the active site of Escherichia coli isocitrate lyase. 总被引:3,自引:1,他引:2 下载免费PDF全文
H G Nimmo F Douglas C Kleanthous D G Campbell C MacKintosh 《The Biochemical journal》1989,261(2):431-435
Escherichia coli isocitrate lyase was inactivated by iodacetate in a pseudo-first-order process. Complete inactivation was associated with the incorporation of only one carboxymethyl group per enzyme subunit. The substrate and products of the enzyme protected against inactivation, suggesting that the reactive group may be located at the active site. Isolation and sequencing of a carboxymethylated peptide showed that the modified residue was a cysteine, in the sequence Cys-Gly-His-Met-Gly-Gly-Lys. The reactivity of isocitrate lyase to iodoacetate declined with pH, following a titration curve for a group of pKa 7.1. The Km of the enzyme for isocritrate declined over the same pH range. 相似文献
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Douglas M. Templeton Anna Paudyn Andrew D. Baines 《Biological trace element research》1989,22(1):17-33
Interest in the biological behavior of a growing number of elements, along with increasing recognition of the importance of interactions among them, demands a versatile and reliable technique for multielement analysis of biological samples. Significant improvements over the sensitivity achieved with conventional inductively coupled plasma (ICP) optical emission spectrometries have been realized with the introduction of quadrupole mass spectrometry (MS) for detection of ions in the plasma. The hybrid technique of ICP-MS promises to be a method of rapid multielement analysis, at detection limits that approach or surpass those of other technologies. However, the application of ICP-MS to analyses of biological interest is truly in its infancy. Here we report the use of ICP-MS for the determination of more than 30 elements of biological interest in a tissue and a biological fluid (rat liver and serum, respectively). Experimental values of the elements serve as a basis for discussion of analytical protocols, performance criteria, and certain problems peculiar to ICP-MS. 相似文献
54.
The glue protein of ribbed mussels (Geukensia demissa): a natural adhesive with some features of collagen 总被引:1,自引:0,他引:1
J. Herbert Waite Douglas C. Hansen Kathleen T. Little 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1989,159(5):517-525
Summary The Atlantic ribbed musselGeukensia (Modiolus)demissa attaches itself to the roots of cord grass and other hard objects in tidal salt marshes by spinning adhesive byssal threads. The precursor of a protein apparently present in the adhesive plaques of the threads was isolated in quantity from the foot of the mussel. The protein has an apparent molecular weight of 130000, a pI of 8.1, and contains a high proportion of Gly, Glu/Gln, Lys and 3,4-dihydroxyphenyl-l-alanine (DOPA). Sequence of tryptic peptides suggests a pattern of repeated motifs, such as: Gly-DOPA-Lys, and X-Gly-DOPA-Y-Z-Gly-DOPA/Tyr-Lys, where X is Thr or Ala in octapeptides and Gln-Thr in nonapeptides. Y is variable, but more often than not hydrophobic; and Z is frequently Pro or 4-trans-hydroxyproline (Hyp). The presence of Pro-Gly and Hyp-Gly sequences of -hydroxylysine in the protein is reminiscent of typical collagens; however, the protein is not labile to clostridial collagenase, nor does collagen cross-react with antibodies raised against the mussel protein. Unlike typical collagens, Gly probably occurs only at every 4th or 5th residue in this unusual mussel protein.Abbreviations
Anti-Gdfp
anti-G. demissa foot protein
-
Dopa
3,4-dihydroxyphenylalanine
-
CTAB
cetyltrimethylammonium bromide
-
HPLC
high performance liquid chromatography
-
PAGE
polyacrylamide gel electrophoresis
Supported by grants from the National Science Foundation (DMB 8500301) and the Office of Naval Research (N00014-86K-0717) 相似文献
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Electroantennogram responses of the Mediterranean fruit fly, Ceratitis capitata to identified volatile constituents from calling males 总被引:1,自引:0,他引:1
Eric B. Jang Douglas M. Light Robert A. Flath Janice T. Nagata Thomas R. Mon 《Entomologia Experimentalis et Applicata》1989,50(1):7-19
Fifty-six compounds from the odor of calling, sexually mature, laboratory reared males of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) were isolated by headspace trapping on Tenax columns and identified using GC/MS techniques (69 total compounds were detected). Electroantennogram responses (EAGs) to 54 of the 56 identified compounds as well as 5 analogs were tested on both sexes. Significant differences between the sexes in their responsiveness were found in 9 of the 54 identified compounds tested. There was no correlation between the amplitude of the EAG response and the relative abundance of compound identified from headspace analysis. Of the five major identified components, three elicited relatively small EAG responses, while two elicited large EAGs compared to the hexan-1-ol standard. The relative ranking of EAG responses were: methyl and ethyl hexenoates and hexanoates > C4–C6 esters and/or acetates > ethyl and methyl octenoates > monoterpenes > sesquiterpenes > C2–C5 acetates, alcohols and ketones. Behavioral bioassays on each of the five major identified components as well as a blend of six of the compounds showed some degree of attractancy to virgin females which in some cases approached the response to a pheromonal standard (male odors absorbed onto filter paper). These results are discussed in relationship to the insect's antennal sensitivity to putative pheromone components and/or allomonal components and to other reported C. capitata pheromone studies.
Résumé Cinquante-six composés de l'odeur de mâles de C. capitata Weidemann, élevés en laboratoire, sexuellement mûrs et en appel, ont été isolés par piégeage sur colonnes tenax et identifiés par la technique GC/MS (69 composés avaient été détectés en tout). Les électroantennogrammes (EAGs) ont été examinés chez les deux sexes pour 54 des 56 composés identifiés et 5 de leurs analogues. Des différences significatives entre les sexes ont été observées pour 9 des 54 composés identifiés. Il n'y avait pas de corrélation entre l'ampleur de l'EAG et l'abondance relative du composé lors de son isolement. Pour les 5 principaux composés identifiés, 3 ont induit des EAGs relativement faibles, tandis que 2 étaient importants, par comparaison avec l'Hexane-1-ol utilisé comme témoin. Le classement relatif des EAG a été: hexénoates et hexanoates d'éthyl et de méthyl C4–C6 esters et/ou acétates octénoates d'éthyl ou de méthyl monoterpènes sesquiterpènes C2–C5 acétates, alcools et kétones. Les expériences de comportement avec chacun des 5 composés principaux identifiés, comme avec des mélanges de 6 composés ont mis en évidence une attraction des femelles vierges qui dans quelques cas avoisine la réponse à la phéromone témoin (odeur du mâle absorbée sur papier filtre). Ces résultats sont discutés en fonction de la sensibilité de l'antenne d'insexte aux composés supposés de la phéromone et aux composés allomonaux, et en fonction des autres études connues sur les phéromones de C. capitata.相似文献
57.
Stereoselective, strong inhibition of ribonucleotide reductase from E. coli by cisplatin 总被引:1,自引:0,他引:1
Using ribonucleotide reductase (EC 1.17.4.1) purified from E. coli clones with overproducing plasmids for the B1 and B2 subunits, respectively, studies have been carried out of the inhibition of this enzyme by cisplatin. Under anaerobic conditions, using the dithiol, reduced form of the enzyme, it was found that ribonucleotide reductase is extremely sensitive to cisplatin: greater than 90% inhibition was achieved with 2-fold molar excess of platinum reagent even at 10(-8)M enzyme. Inhibition was essentially instantaneous and irreversible to G-25 gel filtration. The site of inhibition was found to be the B1 subunit. Transplatin was much less effective. Inhibition of the enzyme by cisplatin (molar ratio cisplatin:B1 = 4.3) led to a decrease in thiol titre corresponding to approximately 1 thiol group per dimer of B1 subunits under conditions leading to 94% inactivation of the ribonucleotide reductase activity. 相似文献
58.
Douglas G. Sprugel 《Trees - Structure and Function》1990,4(2):88-98
Summary Woody-tissue respiration was measured on five different dates at three to five locations on each of 12 30-year-old Abies amabilis trees. On any given date, temperature-corrected respiration per unit surface area varied 10 to 40-fold between sampling locations. In stems, the two major components of respiration were growth respiration and sapwood maintenance respiration, which were of roughly equal importance during the growing season. There was no evidence of significant cambial maintenance respiration, suggesting that a stand with high bole surface area would not automatically have high respiration. Respiration in branches was much greater than in boles of comparable volume and growth rates, and was significantly correlated with branch height. Branch respiration may include an another significant component in addition to the two seen in bole respiration, possibly associated with carbohydrate mobilization and transport or with CO2 efflux from the transpiration stream. 相似文献
59.
β-thalassemia mutations in the Portuguese population 总被引:2,自引:0,他引:2
M. P. Coutinho Gomes M. G. Gomes da Costa L. B. Braga N. T. Cordeiro-Ferreira A. Loi M. Pirastu A. Cao 《Human genetics》1988,78(1):13-15
Summary In this study we have carried out haplotype analysis on the -globin gene cluster and characterized the -thalassemia mutation by oligonucleotide hybridization in 14 patients with thalassemia major and 5 with sickle cell/-thalassemia originating from southern Portugal. We found that three mutations, namely the °-39, ° IVS-1 nt 1 and + IVS-1 nt 110 are prevalent accounting for 53%, 32% and 10% of the -thalassemia chromosomes respectively. In general each mutation was associated with a specific chromosomal haplotype; the ° mutation, however, was linked to three different haplotypes. These results indicate that three oligo-probes complementary to the most common mutations allow prenatal diagnosis by oligonucleotide analysis in 96% of the couples at risk of having offspring with thalassemia major in southern Portugal. 相似文献
60.