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1.
Anastassia Kanavarioti 《Origins of life and evolution of the biosphere》1986,17(1):85-103
We have studied the hydrolysis of guanosine 5-phospho-2-methylimidazolide, 2-MeImpG, in aqueous buffered solutions of various pH's at 75°C and 37°C. At 75°C and pH1.0, two kinetic processes were observed spectrophotometrically: the first and more rapid one is attributed to the hydrolysis of the phosphoimidazolide P-N bond; the second and much slower one, to the cleavage of the glycosidic bond. At 37°C, pH 2.0, the spectrophotometrically determined rate constant of P–N bond hydrolysis was confirmed by using high pressure liquid chromatography, HPLC. With the latter technique it was possible to separate reactants and products and also to extend the pH-rate profile into the neutral region where rates are slower and, therefore, difficult to measure spectrophotometrically. The pH-rate profiles at both temperatures exhibit similar behavior. At pH<2 the pseudo-first-order rate constant increases with decreasing pH; in the region 27. These data are consistent with a reactivity order zwitterion>anion for P–N bond hydrolysis. It is noteworthy that P–N bond hydrolysis in phosphoimidazolides is very slow compared to other phosphoramidates. This may be one of the reasons why this compound showed extraordinary ability in forming long oligomers under template-directed conditions. 相似文献
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I.A. Manuilova L.E. Murashko N.D. Fanchenko V.G. Kolodjko E.A. Chernukha S.A. Rahman M. Bygdeman S.Z. Cekan E. Diczfalusy 《Prostaglandins & other lipid mediators》1979,17(2):313-322
Ten pregnant women (7th–8th week of pregnancy) obtained an intravenous infusion of 15-methyl-prostaglandin-F2α (2.5 μg/min) until clinical signs of abortion occurred or up to 7 hours. Surgical removal of the products of conception was performed 4–7 hours after the start of the infusion. Blood samples were taken prior to and during the infusion and then during the post-abortion period. The plasma levels of both progesterone and estradiol exhibited a significant decrease (p<0.001 and p<0.05, respectively) one hour after the beginning of infusion and remained reduced by approximately 35 and 45 per cent, respectively, during the entire infusion period. A drop in the levels of both steroids was seen after surgical interruption. This was followed by a gradual decrease to non-pregnancy levels. The levels of cortisol increased significantly (p<0.01) by approximately 60 per cent, starting with the second hour of infusion. Following surgical interruption, the levels dropped to pre-infusion values. 17-Hydroxyprogesterone levels increased (p<0.05) above the pretreatment levels by approximately 25 per cent, starting with the third hour of infusion. These levels were not correlated with those of cortisol during the infusion period. Following surgical interruption the plasma levels of 17-hydroxyprogesterone returned to non-pregnancy levels. 相似文献
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Anastassia K. Pogoutse Christine Chieh-Lin Lai Nicholas Ostan Rong-hua Yu Anthony B. Schryvers Trevor F. Moraes 《Analytical biochemistry》2016
Obtaining accurate kinetics and steady-state binding constants for biomolecular interactions normally requires pure and homogeneous protein preparations. Furthermore, in many cases, one of the ligands must be labeled. Over the past decade, several technologies have been introduced that allow for the measurement of kinetics constants for multiple different interactions in parallel. One such technology is bio-layer interferometry (BLI), which has been used to develop systems that can measure up to 96 biomolecular interactions simultaneously. However, despite the ever-increasing throughput of the tools available for measuring protein–protein interactions, the preparation of pure protein still remains a bottleneck in the process of producing high-quality kinetics data. Here, we show that high-quality binding data can be obtained using soluble lysate fractions containing protein that has been biotinylated in vivo using BirA and then applied to BLI sensors without further purification. Furthermore, we show that BirA ligase does not necessarily need to be co-overexpressed with the protein of interest for biotinylation of the biotin acceptor peptide to occur, suggesting that the activity of endogenous BirA in Escherichia coli is sufficient for producing enough biotinylated protein for a binding experiment. 相似文献
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Budde BS Binner P Waldmüller S Höhne W Blankenfeldt W Hassfeld S Brömsen J Dermintzoglou A Wieczorek M May E Kirst E Selignow C Rackebrandt K Müller M Goody RS Vosberg HP Nürnberg P Scheffold T 《PloS one》2007,2(12):e1362
Noncompaction of the ventricular myocardium (NVM) is the morphological hallmark of a rare familial or sporadic unclassified heart disease of heterogeneous origin. NVM results presumably from a congenital developmental error and has been traced back to single point mutations in various genes. The objective of this study was to determine the underlying genetic defect in a large German family suffering from NVM. Twenty four family members were clinically assessed using advanced imaging techniques. For molecular characterization, a genome-wide linkage analysis was undertaken and the disease locus was mapped to chromosome 14ptel-14q12. Subsequently, two genes of the disease interval, MYH6 and MYH7 (encoding the alpha- and beta-myosin heavy chain, respectively) were sequenced, leading to the identification of a previously unknown de novo missense mutation, c.842G>C, in the gene MYH7. The mutation affects a highly conserved amino acid in the myosin subfragment-1 (R281T). In silico simulations suggest that the mutation R281T prevents the formation of a salt bridge between residues R281 and D325, thereby destabilizing the myosin head. The mutation was exclusively present in morphologically affected family members. A few members of the family displayed NVM in combination with other heart defects, such as dislocation of the tricuspid valve (Ebstein's anomaly, EA) and atrial septal defect (ASD). A high degree of clinical variability was observed, ranging from the absence of symptoms in childhood to cardiac death in the third decade of life. The data presented in this report provide first evidence that a mutation in a sarcomeric protein can cause noncompaction of the ventricular myocardium. 相似文献
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Sirpiö S Allahverdiyeva Y Holmström M Khrouchtchova A Haldrup A Battchikova N Aro EM 《The Journal of biological chemistry》2009,284(2):905-912
The NAD(P)H dehydrogenase (NDH) complex functions in photosystem I cyclic electron transfer in higher plant chloroplasts and is crucial for plant responses to environmental stress. Chloroplast NDH complex is a close relative to cyanobacterial NDH-1L complex, and all fifteen subunits so far identified in NDH-1L have homologs in the chloroplast NDH complex. Here we report on the identification of two nuclear-encoded proteins NDH48 and NDH45 in higher plant chloroplasts and show their intimate association with the NDH complex. These two membrane proteins are shown to interact with each other and with the NDH complex enriched in stroma thylakoids. Moreover, the deficiency of either the NDH45 protein or the NDH48 protein in respective mutant plants leads to severe defects in both the accumulation and the function of the NDH complex. The NDH48 and NDH45 proteins are not components of the hydrophilic connecting domain of the NDH complex but are strongly attached to the hydrophobic membrane domain. We conclude that NDH48 and NDH45 are novel nuclear-encoded subunits of the chloroplast NDH complex and crucial both for the stable structure and function of the NDH complex. 相似文献
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