共查询到19条相似文献,搜索用时 62 毫秒
1.
高密度脂蛋白体外氧化修饰动力学研究 总被引:5,自引:1,他引:5
在体外HDL在Cu^2+诱导下可发生氧化修饰,为了探讨体外务浆高密度脂蛋白(HDL)氧化修饰中几种产物的动力学改变,用Cu^2+与HDL保温2 ̄24h,分别观察了HDL氧化修饰过程中硫代巴比妥酸反应物质(TBARS),脂氢过氧化物(LOOH)、共轭二烯(CD)及相对电泳迁移率(REM)等的变化。结果显示,LOOH和CD两个指标动力学变化相似,呈现延滞期,扩增期和下降期三个时相,而TBARS和REM 相似文献
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以健康的中华蟾蜍为实验动物,研究皮肤染毒巯基乙酸(TGA)对血液天门冬氨酸氨基转氨酶(AST)、丙氨酸氨基转氨酶(ALT)和钙含量的影响,并对其作用机理进行初步探讨.试验结果表明,染毒巯基乙酸剂量对于血清AST、ALT和Ca含量影响效果明显.方差分析表明各试验处理组与对照组相比差异显著;染毒TGA的时间对于血清生化指标的影响呈现累积趋势,各试验处理组染毒后期与前期差异显著.表明巯基乙酸对于蟾蜍肝组织等器官具有明显的损伤作用,因此化工产品巯基乙酸对水体的污染不容忽视. 相似文献
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实验用Ms培养基,利用去除铁离子的氧化亚铁硫杆菌(Thiobacillus ferrooxidans)进行了细菌亚硫酸盐的生长代谢研究。实验结果表明氧化亚铁硫杆菌对亚硫酸根具有一定的氧化能力。用Origin 7.0对实验数据进行拟合处理,表明了氧化亚铁硫杆菌催化氧化亚硫酸盐的动力学方程符合Hill方程。氧化亚铁硫杆菌催化氧化亚硫酸盐是一个底物抑制的细胞反应,其KS值随pH值和底物浓度的改变而变化。pH值对反应有很大的影响,pH值越接近中性KS就越小,反应速率就越大。 相似文献
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HcNPV半胱氨酸蛋白酶基因的核苷酸序列研究 总被引:6,自引:1,他引:6
HcNPV半胱氨酸蛋白酶基因的核苷酸序列研究贡成良1小林淳宫岛成寿金伟1吴祥甫2*(日本三重大学分子素材工学科,三重514,日本;1浙江农业大学蚕学系,杭州310029;2中国科学院上海生物化学研究所,上海200031)关键词美国白蛾;核型多角体病毒... 相似文献
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电渗析法分离提纯N-乙酰-L-半胱氨酸研究 总被引:1,自引:1,他引:1
使用我校由辐射法制备的高性能离子交换膜HF—1及HF—2,采用电渗析技术对合成所得的N-乙酰-L-半胱氨酸进行了分离提纯,脱盐率>15%,损失率<15%,为工业化应用提供了依据。 相似文献
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巯基乙酸对中华蟾蜍红细胞核异常的影响 总被引:1,自引:1,他引:1
试验以健康的中华蟾蜍为实验动物,研究注射巯基乙酸对红细胞核异常的影响.试验结果表明,在一定的范围内,巯基乙酸可引起蟾蜍微核细胞率和核异常细胞率遗传指标发生明显变化,随着巯基乙酸浓度的增加和作用时间的延长,蟾蜍红细胞微核和核异常率呈现先上升后下降的规律性变化. 相似文献
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目的:探讨南部战区某旅歼击飞行员血清皮质醇(COR)和同型半胱氨酸(Hcy)联合检测的意义。方法:对歼击飞行员(A组)、地方健康体检人员(B组)、部队考学战士(C组)、空军地勤人员(C组)的血清COR、Hcy水平进行两两比较,通过Kruskal-Wallis统计学方法处理分析。结果:B组人员COR水平比A组、C组、D组人员低,差异有统计学意义(P<0.05);C组人员Hcy水平比A组、B组、D组人员高,差异有统计学意义(P<0.05)。结论:经常性的飞行训练可增强飞行员分泌激素的适应性,在航空保健工作中,COR联合Hcy监测及动态观察对保障飞行员的健康和飞行安全,提高部队战斗力具有一定的意义。 相似文献
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目的:探讨冠心病患者血浆中同型半胱氨酸、内皮素、循环内皮细胞水平与冠状动脉粥样硬化性心脏病发生过程的关系。方法:对62例冠心病患者组和50例健康对照组分别采用高效液相色谱分析法和放射免疫分析法测定血浆中同型半胱氨酸水平和内皮素水平,同时测定血浆中循环内皮细胞的数量。结果:冠心病组同型半胱氨酸(21.27±5.73)μmol/L、内皮素(84.30±13.68)ng/L、循环内皮细胞(12.08±5.85)和cells/0.9μl明显高于对照组同型半胱氨酸(9.12±4.87)μmol/L、内皮素(47.65±12.71)ng/L、循环内皮细胞(2.35±1.02)cells/0.9μl,P均小于0.001。同型半胱氨酸与内皮素、循环内皮细胞呈正相关(r=0.601,P0.01;r=0.645,P0.01),且冠心病组血浆循环内皮细胞和内皮素呈正相关(r=0.850,P0.01)。结论:同型半胱氨酸及内皮素对冠状动脉粥样硬化性心脏病的发生发展有促进作用,对其检测有临床实用价值。 相似文献
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The partial genomic library of Acetobacter suboxydans was constructed using Yeast| E.coli shuttle plasmid YEp352 as vector.Two positive transformants,designated as DH5α(pAD91) and DH5α(pAD98),were obtained by screening the growth of transformants on the agar plate in which D|arabitol was used as the sole carbon source.The results of Southern blot and restriction endonuclease analysis showed that the two recombinants are identical.The insert is about 2.3kb.Arabitol dehydrogenase activity assay indic… 相似文献
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本试验利用生化检测方法,以健康的中华蟾蜍为实验动物,研究注射巯基乙酸对血液生理指标的影响,并对其作用机理进行了初步探讨.试验结果表明,随染毒TGA剂量增加,蟾蜍血清总蛋白、葡萄糖含量下降,胆固醇、甘油三酯含量上升;随染毒TGA时间的延长,血清总蛋白、葡萄糖含量先下降后上升但均低于对照组,胆固醇、甘油三酯含量先上升后下降但均高于对照组.因此提示化工产品对生物的影响不容忽视. 相似文献
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Murakami T Nojiri M Nakayama H Odaka M Yohda M Dohmae N Takio K Nagamune T Endo I 《Protein science : a publication of the Protein Society》2000,9(5):1024-1030
Nitrile hydratase from Rhodococcus sp. N-771 is an alphabeta heterodimer with a nonheme ferric iron in the catalytic center. In the catalytic center, alphaCys112 and alphaCys114 are modified to a cysteine sulfinic acid (Cys-SO2H) and a cysteine sulfenic acid (Cys-SOH), respectively. To understand the function and the biogenic mechanism of these modified residues, we reconstituted the nitrile hydratase from recombinant unmodified subunits. The alphabeta complex reconstituted under argon exhibited no activity. However, it gradually gained the enzymatic activity through aerobic incubation. ESI-LC/MS analysis showed that the anaerobically reconstituted alphabeta complex did not have the modification of alphaCys112-SO2H and aerobic incubation induced the modification. The activity of the reconstituted alphabeta complex correlated with the amount of alphaCys112-SO2H. Furthermore, ESI-LC/MS analyses of the tryptic digest of the reconstituted complex, removed of ferric iron at low pH and carboxamidomethylated without reduction, suggested that alphaCys114 is modified to Cys-SOH together with the sulfinic acid modification of alphaCys112. These results suggest that alphaCys112 and alphaCys114 are spontaneously oxidized to Cys-SO2H and Cys-SOH, respectively, and alphaCys112-SO2H is responsible for the catalytic activity solely or in combination with alphaCys114-SOH. 相似文献
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Yuan‐Chang Chang Chien‐Ning Huang Chia‐Hung Lin Huan‐Cheng Chang Chih‐Che Wu 《Proteomics》2010,10(16):2961-2971
Oxidation of thiol proteins, which results in conversion of cysteine residues to cysteine sulfenic, sulfinic or sulfonic acids, is an important posttranslational control of protein function in cells. To facilitate the analysis of this process with MALDI‐MS, we have developed a method for selective enrichment and identification of peptides containing cysteine sulfonic acid (sulfopeptides) in tryptic digests of proteins based on ionic affinity capture using polyarginine‐coated nanodiamonds as high‐affinity probes. The method was applied to selectively concentrate sulfopeptides from either a highly dilute solution or a complex peptide mixture in which the abundance of the sulfonated analyte is as low as 0.02%. The polyarginine‐coated probes exhibit a higher affinity for peptides containing multiple sulfonic acids than peptides containing single sulfonic acid. The limit of the detection is in the femtomole range, with the MALDI‐TOF mass spectrometer operating in the negative ion mode. The results show that the new approach has good specificity even in the presence of phosphopeptides. An application of this method for selective enrichment and structural identification of sulfopeptides is demonstrated with the tryptic digests of performic‐acid‐oxidized BSA. 相似文献
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B-1625 FA2β, one of the minor acidic actinomycin congeners produced by Streptomyces antibioticus No. B-1625, was produced selectively on the combined use of sarcosine and ferrous sulfate in a complex medium. The FA2β was further separated into two components, MeOH- insoluble FA2β – 1 and MeOH-soluble FA2β – 2, by methanol treatment. After repeating the MeOH treatment, the purified FA2β – 1 was examined as to its physicochemical properties, and subjected to amino acid analysis, and 13C-, 1H-NMR and FAB mass spectral measurements. As a result, the structure of FA2β – 1 was deduced to be 2-amino-1-carboxyl-4,6-dimethyl-3-phenoxazone-9- carbonyl-threonyl-valyl-sarcosyl-sarcosyl-N-methylvaline. As much as 1 mg/ml of FA2β – 1 showed no antibacterial activity against Gram-negative or Gram-positive bacteria, although FA2β – 2 shows antibiotic activity. 相似文献
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One of the most popular and simple models for the calculation of pKas from a protein structure is the semi‐macroscopic electrostatic model MEAD. This model requires empirical parameters for each residue to calculate pKas. Analysis of current, widely used empirical parameters for cysteine residues showed that they did not reproduce expected cysteine pKas; thus, we set out to identify parameters consistent with the CHARMM27 force field that capture both the behavior of typical cysteines in proteins and the behavior of cysteines which have perturbed pKas. The new parameters were validated in three ways: (1) calculation across a large set of typical cysteines in proteins (where the calculations are expected to reproduce expected ensemble behavior); (2) calculation across a set of perturbed cysteines in proteins (where the calculations are expected to reproduce the shifted ensemble behavior); and (3) comparison to experimentally determined pKa values (where the calculation should reproduce the pKa within experimental error). Both the general behavior of cysteines in proteins and the perturbed pKa in some proteins can be predicted reasonably well using the newly determined empirical parameters within the MEAD model for protein electrostatics. This study provides the first general analysis of the electrostatics of cysteines in proteins, with specific attention paid to capturing both the behavior of typical cysteines in a protein and the behavior of cysteines whose pKa should be shifted, and validation of force field parameters for cysteine residues. Proteins 2012. © 2012 Wiley Periodicals, Inc. 相似文献
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The aim of this study was to design and evaluate of mucoadhesive gel formulations for the vaginal application of clomiphene
citrate (CLM) for local treatment of human papilloma virus (HPV) infections. Chitosan (CHI) and polycarbophil (PC) were covalently
modified using the thioglycolic acid and L-cysteine, respectively. The formation of thiol conjugates of chitosan (CHI-TG)
and polycarbophil (PC-CYS) were confirmed by FT-IR analysis and PC-CYS and CHI-TG were found to have 148.42 ± 4.16 and 41.17 ± 2.34 μmol
of thiol groups per gram of polymer, respectively. One percent CLM gels were prepared by combination of various concentrations
of PC and CHI with thiolated conjugates of these polymers. Hardness, compressibility, elasticity, adhesiveness and cohesiveness
of the gels were measured by Texture profile analysis and the vaginal mucoadhesion was investigated by mucoadhesion test.
The increasing in the amount of the thiol conjugates was found to enhance the elasticity, cohesiveness, adhesiveness and mucoadhesion
of the gel formulations but not their hardness and compressibility when compared to gels prepared using their respective parent
formulations. Slower release rate of CLM from gels was achieved when the polymer concentrations were increased in the gel
formulations. PC and its thiol conjugate were found to prolong the release of CLM longer than 70 h unlike gel formulations
prepared using CHI and its thiol conjugate which were able to release CLM up to 12 h. Stability of CLM was preserved during
the 3 month stability analysis under controlled room temperature and accelerated conditions. 相似文献
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The triethylammonium QX-314 and the trimethylammonium QX-222 are lidocaine derivatives that act as open-channel blockers of the acetylcholine (ACh) receptor. When bound, these blockers should occlude some of the residues lining the channel. Eight residues in the second membrane-spanning segment (M2) of the mouse-muscle α subunit were mutated one at a time to cysteine and expressed together with wild-type β, γ, and δ subunits in Xenopus oocytes. The rate constant for the reaction of each substituted cysteine with 2-aminoethyl methanethiosulfonate (MTSEA) was determined from the time course of the irreversible effect of MTSEA on the ACh-induced current. The reactions were carried out in the presence and absence of ACh and in the presence and absence of QX-314 and QX-222. These blockers had no effect on the reactions in the absence of ACh. In the presence of ACh, both blockers retarded the reaction of extracellularly applied MTSEA with cysteine substituted for residues from αVal255, one third of the distance in from the extracellular end of M2, to αGlu241, flanking the intracellular end of M2, but not with cysteine substituted for αLeu258 or αGlu262, at the extracellular end of M2. The reactions of MTSEA with cysteines substituted for αLeu258 and αGlu262 were considerably faster in the presence of ACh than in its absence. That QX-314 and QX-222 did not protect αL258C and αE262C against reaction with MTSEA in the presence of ACh implies that protection of the other residues was due to occlusion of the channel and not to the promotion of a less reactive state from a remote site. Given the 12-Å overall length of the blockers and the α-helical conformation of M2 in the open state, the binding site for both blockers extends from αVal255 down to αSer248. 相似文献
19.
Pan JC Cheng Y Hui EF Zhou HM 《Biochemical and biophysical research communications》2004,317(2):539-544
The reduction of 5,5'-dithiobis-(2-nitrobenzoic acid)-modified arginine kinase by dithiothreitol has been investigated using the kinetic theory of the substrate reaction during modification of enzyme activity. The results show that the modified arginine kinase can be fully reactivated by an excess concentration of dithiothreitol in a monophasic kinetic course. The presence of ATP or the transition-state analog markedly slows the apparent reactivation rate constant, while arginine shows no effect. The results of ultraviolet (UV) difference and intrinsic fluorescence spectra indicate that the substrate arginine-ADP-Mg2+ can induce conformational changes of the modified enzyme but adding NO3- cannot induce further changes that occur with the native enzyme. The reactive cysteines' location and role in the catalysis of arginine kinase are discussed. It is suggested that the cysteine may be located in the hinge region of the two domains of arginine kinase. The reactive cysteine of arginine kinase may play an important role not in the binding to the transition-state analog but in the conformational changes caused by the transition-state analog. 相似文献