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11.
F F Kadlubar K C Morton D M Ziegler 《Biochemical and biophysical research communications》1973,54(4):1255-1261
Organic hydroperoxides are capable of supporting the C-oxidation of several different amines in the presence of hepatic microsomes. Evidence is presented that indicates that microsomal cytochrome P-450 acts as the catalyst. Removal of the NADPH-cytochrome oxidoreductase or essential phospholipid from microsomes does not significantly affect the peroxidase activity. Of the amine substrates C-oxidized by organic hydroperoxides in the presence of microsomes, only aminopyrine and dimethylaniline are rapidly oxidized by hydroperoxides in the presence of catalase. The catalase-mediated reaction can also be distinguished from the microsomal-catalyzed reaction by the use of differential inhibitors. 相似文献
12.
Gordon C. K. Roberts 《Neurochemical research》1996,21(9):1117-1124
NMR spectroscopy has proved to be a valuable tool in the study of the interactions between enzymes and their substrates. The
kinds of structural and dynamic information which can be obtained are illustrated by studies of three enzymes involved in
drug metabolism. Cytochromes P450 play a crucial role in metabolism of a wide range of exogenous chemicals. NMR has been used to measure distances from the
haem iron of the cytochrome to protons of the bound substrate, leading to detailed structural models for the enzyme-substrate
complexes. The other two enzymes, chloramphenicol acetyltransferase and β-lactamase, are responsible for bacterial resistance
to specific antibiotics. In chloramphenicol acetyltransferase, NMR has been used to determine the conformation of coenzyme
A bound to the enzyme, while in the case of β-lactamase the pK of a specific lysine residue at the active site has been determined,
providing valuable information on the catalytic mechanism.
Special issue dedicated to Dr. Herman Bachelard. 相似文献
13.
Francisco M. Pinto Yves Chupeau Vicente M. Cabrera 《In vitro cellular & developmental biology. Plant》1995,31(2):96-100
An efficient and easy method for genetic characterization of plant somatic hybrids is proposed. In a first qualitative approach,
four somatic hybrids and their parental species (Nicotiana tabacum andN. plumbaginifolia) were characterized by DNA fingerprinting and Random Amplification of Polymorphic DNA (RAPD). After this, a quantitative
estimation of the degree of parental contribution to the hybrids was carried out by means of a slot-blot analysis. Both qualitative
methods, showed one hybrid identical toN. tabacum, two almost identical toN. plumbaginifolia, and a fourth similar to this parental species, but with someN. tabacum admixture. The quantitative method, for the same hybrids, gave 83%, 7%, 7%, and 37%N. tabacum DNA contribution, respectively. 相似文献
14.
Na2SO3对热-DTT活化的游离CF1及类囊体膜上CF1-ATPase活力均有显著的促进作用,NaHCO3亦有明显的促进作用。Na2SO3和NaHCO3的促进作用与它们解除Mg2+的抑制作用有关。从NaHCO3和Na2SO3及它们与Mg2+之间的竞争性关系,表明三者是结合在酶的同一部位上。Na2SO3可明显降低热-DTT活化的游禹CF1-ATPase催化反应的活化能,这可能与促进产物ADP的释放有关。 相似文献
15.
Resistance to potato leafroll virus (PLRV), potato virus Y (PVYo) and potato virus X (PVX) was studied in symmetric and asymmetric somatic hybrids produced by electrofusion between Solanum brevidens (2n=2×=24) and dihaploid S. tuberosum (2n=2×=24), and also in regenerants (B-hybrids) derived through protoplast culture from a single somatic hybrid (chromosome number 48). All of the somatic hybrids between 5. brevidens and the two dihaploid lines of potato cv. Pito were extremely resistant to PLRV and PVYoand moderately resistant to PVX, irrespective of their chromosome number and ploidy level (tetraploid or hexaploid). Most (56%) of the asymmetric hybrids of irradiated S. brevidens and the dihaploid line of potato cv. Pentland Crown (PDH40) had high titres of PVYosimilar to those of PDH40, whereas the rest of the hybrids had PVYotitres less than a tenth of those in PDH40. Three B-hybrids had a highly reduced chromosome number (27, 30 and 34), but were however as resistant to PLRV, PVYoand PVX as 5. brevidens. Two asymmetric hybrids and one B-hybrid were extremely resistant to PLRV but susceptible to both PVY and PVX. The results suggested that resistance to PLRV in 5. brevidens is controlled by a gene or genes different from those controlling resistance to PVY and PVX, and the gene(s) for resistance to PVY and PVX are linked in S. brevidens. 相似文献
16.
A 70-amino acid zinc-binding polypeptide fragment from the regulatory chain of aspartate transcarbamoylase causes marked changes in the kinetic mechanism of the catalytic trimer.
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B. B. Zhou G. L. Waldrop L. Lum H. K. Schachman 《Protein science : a publication of the Protein Society》1994,3(6):967-974
Interaction between a 70-amino acid and zinc-binding polypeptide from the regulatory chain and the catalytic (C) trimer of aspartate transcarbamoylase (ATCase) leads to dramatic changes in enzyme activity and affinity for active site ligands. The hypothesis that the complex between a C trimer and 3 polypeptide fragments (zinc domain) is an analog of R state ATCase has been examined by steady-state kinetics, heavy-atom isotope effects, and isotope trapping experiments. Inhibition by the bisubstrate ligand, N-(phosphonacetyl)-L-aspartate (PALA), or the substrate analog, succinate, at varying concentrations of substrates, aspartate, or carbamoyl phosphate indicated a compulsory ordered kinetic mechanism with carbamoyl phosphate binding prior to aspartate. In contrast, inhibition studies on C trimer were consistent with a preferred order mechanism. Similarly, 13C kinetic isotope effects in carbamoyl phosphate at infinite aspartate indicated a partially random kinetic mechanism for C trimer, whereas results for the complex of C trimer and zinc domain were consistent with a compulsory ordered mechanism of substrate binding. The dependence of isotope effect on aspartate concentration observed for the Zn domain-C trimer complex was similar to that obtained earlier for intact ATCase. Isotope trapping experiments showed that the compulsory ordered mechanism for the complex was attributable to increased "stickiness" of carbamoyl phosphate to the Zn domain-C trimer complex as compared to C trimer alone. The rate of dissociation of carbamoyl phosphate from the Zn domain-C trimer complex was about 10(-2) that from C trimer.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
17.
Arne O. Smals Eldbjrg S. Heimstad Asbjrn Hordvik Nils P. Willassen Rune Male 《Proteins》1994,20(2):149-166
The crystal structure of an anionic form of salmon trypsin has been determined at 1.82 Å resolution. We report the first structure of a trypsin from a phoikilothermic organism in a detailed comparison to mammalian trypsins in order to look for structural rationalizations for the cold-adaption features of salmon trypsin. This form of salmon trypsin (T II) comprises 222 residues, and is homologous to bovine trypsin (BT) in about 65% of the primary structure. The tertiary structures are similar, with an overall displacement in main chain atomic positions between salmon trypsin and various crystal structures of bovine trypsin of about 0.8 Å. Intramolecular hydrogen bonds and hydrophobic interactions are compared and discussed in order to estimate possible differences in molecular flexibility which might explain the higher catalytic efficiency and lower thermostability of salmon trypsin compared to bovine trypsin. No overall differences in intramolecular interactions are detected between the two structures, but there are differences in certain regions of the structures which may explain some of the observed differences in physical properties. The distribution of charged residues is different in the two trypsins, and the impact this might have on substrate affinity has been discussed. © 1994 Wiley-Liss, Inc. 相似文献
18.
非同位素PCR-单链构象多态性技术的建立和应用 总被引:2,自引:0,他引:2
PCR-单链构象多态性技术问世以来,成为研究基因突变的工具,特别是在分子肿瘤学研究中,广泛应用于癌基因,抑癌基因突变的研究,常规PCR-SSCP采用同位素标记PCR产物,测序板电泳分离突变,在操作和费用上有种种局限,文章建立了一种非同位素PCR-SSCP技术;通过不对称PCR获得单链,普通PAGE分离,经银染检出突变,用这种方法,还研究了四株鼻咽癌细胞株CNE1,CNE2,HK1和SUNE1中肿瘤 相似文献
19.
Trees at their upper range limits are highly sensitive to climate change, and thus alpine treelines worldwide have changed their recruitment patterns in response to climate warming. However, previous studies focused only on daily mean temperature, neglecting the asymmetric influences of daytime and nighttime warming on recruitments in alpine treelines. Here, based on the compiled dataset of tree recruitment series from 172 alpine treelines across the Northern Hemisphere, we quantified and compared the different effects of daytime and nighttime warming on treeline recruitment using four indices of temperature sensitivity, and assessed the responses of treeline recruitment to warming-induced drought stress. Our analyses demonstrated that even in different environmental regions, both daytime and nighttime warming could significantly promote treeline recruitment, and however, treeline recruitment was much more sensitive to nighttime warming than to daytime warming, which could be attributable to the presence of drought stress. The increasing drought stress primarily driven by daytime warming rather than by nighttime warming would likely constrain the responses of treeline recruitment to daytime warming. Our findings provided compelling evidence that nighttime warming rather than daytime warming could play a primary role in promoting the recruitment in alpine treelines, which was related to the daytime warming-induced drought stress. Thus, daytime and nighttime warming should be considered separately to improve future projections of global change impacts across alpine ecosystems. 相似文献
20.
Synthetic chimeras of mouse growth factor-associated glandular kallikreins. I. Kinetic properties. 总被引:1,自引:1,他引:0
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M. Blaber P. J. Isackson J. P. Burnier J. C. Marsters Jr R. A. Bradshaw 《Protein science : a publication of the Protein Society》1993,2(8):1210-1219
A series of six chimeric proteins, composed of fragments corresponding to either one or the other of the growth factor-associated mouse glandular kallikreins-epidermal growth factor binding protein (EGF-BP) and the gamma-subunit of nerve growth factor (gamma-NGF)--were expressed in Escherichia coli and isolated, and their kinetic properties were characterized. The assembly of these synthetic proteases involved the substitution of regions of the proteins containing four specific surface loops that have been postulated to influence both kinetic specificity and the formation of growth factor complexes. The substrates utilized in the kinetic characterization of these chimeric kallikreins were tripeptide nitroanilides representing carboxyl termini of both the EGF and beta-NGF mature hormones, putative processing sites for these kallikreins in the precursors. Characterization of these hybrid enzymes demonstrates that Km and kcat kinetic constants may be independently affected by the regions utilized in construction of these chimeric kallikreins. Specifically, loop 1, located in the amino terminal region (Bode, W., et al., J. Mol. Biol. 164, 237-282, 1983), in gamma-NGF enhanced the kcat for substrates containing threonine in the P2 position, as is the case during the processing of the carboxy terminus of the beta-NGF precursor. Also, the central regions of the kallikreins containing loop 2 and the kallikrein loop dictated the generally inverted Km and kcat kinetic constants observed between EGF-BP and gamma-NGF. Finally, in gamma-NGF the autolysis loop, found in the carboxyl terminal region, functions to lower the Km kinetic constant for a variety of substrates. The results allow previously characterized kinetic differences between EGF-BP and gamma-NGF to be interpreted in terms of specific regions of the proteins and identify a subset of amino acid positions responsible for these functional characteristics. 相似文献