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31.
A preparation of ATPase from the membranes of Micrococcus lysodeikticus, solubilized and more than 95 %. pure, showed two main bands in analytical polyacrylamide gel electrophoresis. They did not correspond to isoenzymes because one band could be converted into the other by exposure to a mildly alkaline pH value. The conversion was paralleled by changes in molecular weight, circular dichroism and catalytic properties. Denaturation by pH at 25 °C was followed by means of circular dichroism, ultracentrifugation and polyacrylamide gel electrophoresis. A large conformational transition took place in the acid range with midpoints at about pH = 3.6 (I = 10?4 M), 4.3 (I = 0.03 M) and 5.3 (I = 0.1 M). The transition was irreversible. Strong aggregation of the protein occurred in this range of pH. The final product was largely random coil, but even at pH 1.5 dissociation into individual subunits was not complete. However, partial dissociation took place at pH 5 (I = 0.028 M). At this pH value the enzyme was inactive, but 20–30 % of the activity could be recovered when the pH was returned to 7.5.In the alkaline region the midpoint of the transition occurred near pH = 11 (I = 0.028 M). The pK of most of the tyrosine residues of the protein was about 10.9. The unfolding was irreversible and the protein was soon converted into peptide species with molecular weights lower than those determined for the subunits by gel clectrophoresis in the presence of sodium dodecyl sulphate. Conventional proteolysis did not account for the transformation. 相似文献
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33.
Irradiating the mycelium of Neurospora crassa with moderate intensities of blue light causes a reversible photoreduction of a b-type cytochrome. The action spectrum for the photoreduction of cytochrome b is very similar to the absorption spectrum of flavin pigments. Prolonged irradiation of the mycelium with strong blue light irreversibly bleaches flavin-like pigments and as these pigments are bleached the photoresponse of cytochrome b is lost. We conclude from these and other data that a flavin is the photoreceptor pigment for the photoreduction of cytochrome b. The close similarity between the action spectrum for the photoreduction of cytochrome b and action spectra for a number of physiological photoresponses suggests that this photoreceptor pigment controls a wide variety of photobiological processes in a wide diversity of organisms. 相似文献
34.
B A Faraj J Y Mu M S Lewis J P Wilson Z H Israili P G Dayton 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1975,149(3):664-669
Antibodies were prepared against tyramine. The antigen was prepared as follows: p-Aminohippuric acid was coupled to mBSA using a carbodiimide reagent. The amino group was diazotized an attached to the aromatif ring of TYR. The immunogen in Freund's complete adjuvant was injected into rabbits. The specificity of the resulting antibody was determined by radioimmunoassay. Using random-labeled TYR-3H, TYR, its metabolites, phenethylamine analogs, catecholamines, and certain amino acids were evaluated by a competitive binding assay method. With this technique 4 ng of TYR inhibited the binding of TYR-3H by 50%. The radioimmunoassay of TYR was used to measure the plasma, urine, and tissue levels of TYR in rabbits. The plasma disappearance curve of TYR revealed a biphasic pattern with t1/2 of 2 min and 54 min. The highest concentration of TYR was found in adrenals and spleen. The factthat the major metabolites of TYR and a series of pharmacologically important sympathomimetics and catecholamines did not interfere, makes the radioimmunoassay of TYR a useful, simple, sensitive, and spedific method for the direct analysis of TYR in biological meterials. 相似文献
35.
【背景】接合菌在自然界广泛分布,在工业、食品、医药、生物防治等方面扮演着重要的角色,部分接合菌为有害菌。西藏地区接合菌只有少数零星的报道,其系统调查研究几乎还是空白,大量潜在的物种需要分离、鉴定、认识、保藏和开发。【目的】了解西藏地区的接合菌物种多样性和生物资源现状,为该地区有害接合菌的控制和有益接合菌的开发利用奠定基础。【方法】对西藏全境7个地区19个代表县的701个样品采用平板稀释分离法得到单菌落,然后进行形态和分子(SSU、ITS和LSU r DNA)鉴定,并在此基础上进行生物多样性以及优势和稀有属种分析。【结果】得到西藏接合菌10属26种,包括5个西藏已知种和21个西藏新记录种;其中4个为中国新记录种,分别是:类变形被孢霉(Mortierella amoeboidea)、球孢高山被孢霉(M.globalpina)、灰褐毛霉(Mucor brunneogriseus)、暗色毛霉(M.fuscus)。西藏接合菌的多样性指数分析表明,不同地区间的物种数量和组成存在显著差异,排在前4位的分别是波密县、米林县、当雄县和八宿县。属种分析显示,西藏接合菌的2个优势属为被孢霉属(Mortierella)和毛霉属(Mucor);3个优势种为高山被孢霉(Mortierella alpina)、冻土毛霉(Mucor hiemalis)、匍枝根霉(Rhizopus stolonifer);常见属8个,分别是犁头霉属(Absidia)、放射毛霉属(Actinomucor)、小克银汉霉属(Cunninghamella)、根毛霉属(Rhizomucor)、根霉属(Rhizopus)、共头霉属(Syncephalastrum)、伞形霉属(Umbelopsis)和接霉属(Zygorhynchus);常见种9个,稀有种14个。【结论】西藏地区接合菌资源丰富,各地区生物多样性差异显著,稀有物种占一半以上的比例提示西藏自然环境保护的重要性。 相似文献
36.
The NMR determination of the structure of Cd(7)-metallothionein was done previously using a relatively large protein concentration that favors dimer formation. The reactivity of the protein is also affected under this condition. To examine the influence of protein concentration on metallothionein conformation, the isolated Cd(4)-alpha-domain was prepared from rabbit metallothionein-2 (MT 2), and its three-dimensional structure was determined by heteronuclear, (1)H-(111)Cd, and homonuclear, (1)H-(1)H NMR, correlation experiments. The three-dimensional structure was refined using distance and angle constraints derived from these two-dimensional NMR data sets and a distance geometry/simulated annealing protocol. The backbone superposition of the alpha-domain from rabbit holoprotein Cd(7)-MT 2 and the isolated rabbit Cd(4)-alpha was measured at a RMSD of 2.0 A. Nevertheless, the conformations of the two Cd-thiolate clusters were distinctly different at two of the cadmium centers. In addition, solvent access to the sulfhydryl ligands of the isolated Cd(4)-alpha cluster was 130% larger due to this small change in cluster geometry. To probe whether these differences were an artifact of the structure calculation, the Cd(4)-alpha-domain structure in rabbit Cd(7)-MT 2 was redetermined, using the previously defined set of NOEs and the present calculation protocol. All calculations employed the same ionic radius for Cd(2+) and same cadmium-thiolate bond distance. The newly calculated structure matched the original with an RMSD of 1.24 A. It is hypothesized that differences in the two alpha-domain structures result from a perturbation of the holoprotein structure because of head-to-tail dimerization under the conditions of the NMR experiments. 相似文献
37.
分离自冬虫夏草可培养真菌的多样性研究 总被引:2,自引:1,他引:2
冬虫夏草是生长于青藏高原的一种名贵中药材。天然冬虫夏草及其微环境中生活着多种真菌。作者使用常规分离培养方法对冬虫夏草的真菌区系进行研究。从天然冬虫夏草的子座、菌核和菌膜3个部位共分离到572个真菌菌株,并根据形态特征将大部分菌株鉴定到37个不同的属。这些菌株经SSCP(single-strand conformation polymorphism)分析后,再根据nrDNAITS序列的相似性(以97%为阈值)共区分出92种不同的分类单元(operational taxonomic unit,OTU)。其中,属于子囊菌的菌株数及OTU数均比接合菌和担子菌多。从菌膜分离的菌株数及OTU数都明显多于子座和菌核。分离自子座的优势真菌是产黄青霉Penicillium chrysogenum,而分离自菌核和菌膜的优势真菌均为玫红假裸囊菌Pseudogymnoascus roseus。尚未最终鉴定的部分真菌可能为新的真菌物种。 相似文献
38.
Recently, a class of about 22 nucleotides (nt) small RNA has been discovered in many eukaryotes, termed microRNAs (miRNAs),
which have a variety of functions. Many recent findings have demonstrated that viruses can also encode their own miRNAs. Meanwhile,
other findings reveal a relationship between host miRNA and viral infection. These findings suggest a tight relationship between
host and viral infection via miRNA pathway. This article introduces the miRNAs encoded by viruses and reviews the advances
of the interaction of the mammalian host miRNAs and viral infection. 相似文献
39.
40.
Zhang K Zang TZ Yang W Sun Y Mu Y Liu JQ Shen JC Luo GM 《The Journal of biological chemistry》2006,281(18):12516-12520
Substrate binding and the subsequent reaction are the two principal phenomena that underlie the activity of enzymes, and many enzyme-like catalysts were generated based on the phenomena. The single chain variable region fragment of antibody 2F3 (scFv2F3) was elicited against hapten GSH-S-DN2phBu, a conjugate of glutathione (GSH), butyl alcohol, and 1-chloro-2,4-dinitrobenzene (CDNB); it can therefore bind both GSH and CDNB, the substrates of native glutathione S-transferases (GSTs). It was shown previously that there is a serine residue that is the catalytic group of GST in the CDR regions of scFv2F3 close to the sulfhydryl of GSH. Thus, we anticipated that scFv2F3 will display GST activity. The experimental results showed that scFv2F3 indeed displayed GST activity that is equivalent to the rat-class GST T-2-2 and exhibited pH- and temperature-dependent catalytic activity. Steady-state kinetic studies showed that the Km values for the substrates are close to those of native GSTs, indicating that scFv2F3 has strong affinities for the substrates. Compared with some other GSTs, its kcat value was found to be low, which could be caused by the similarity between the GSH-S-DN2phBu and the reaction product of GSH and CDNB. These results showed that our approach to imitating enzymes is correct, which is that an active site may catalyze a chemical reaction when a catalytic group locates beside a substrate-binding site of a receptor. It is important to consider product inhibition in hapten design in order to obtain a mimic with a high catalytic efficiency. 相似文献