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101.
The three-dimensional structures of two isozymes of manganese peroxidase (MnP) have been predicted from homology modeling using lignin peroxidase as a template. Although highly homologous, MnP differs from LiP by the requirement of Mn(II) as an intermediate in its oxidation of substrates. The Mn(II) site is absent in LiP and unique to the MnP family of peroxidases. The model structures were used to identify the unique Mn(II) binding sites, to determine to what extent they were conserved in the two isozymes, and to provide insight into why this site is absent in LiP. For each isozyme of MnP, three candidate Mn(II) binding sites were identified. Energy optimizations of the three possible Mn(II) enzyme complexes allowed the selection of the most favorable Mn(II) binding site as one with the most anionic oxygen moieties best configured to act as ligands for the Mn(II). At the preferred site, the Mn(II) is coordinated to the carboxyl oxygens of Glu-35, Glu-39, and Asp-179, and a propionate group of the heme. The predicted Mn(II) binding site is conserved in both isozymes. Comparison between the residues at this site in MnP and the corresponding residues in LiP shows that two of the three anionic residues in MnP are replaced by neutral residues in LiP, explaining why LiP does not bind Mn(II). © 1994 Wiley-Liss, Inc. 相似文献
102.
Inorganic cofactors (Mn, Ca2+ and Cl-) are essential for oxidation of H2O to O2 by Photosystem II. The Mn reductants NH2OH and its N-methyl derivatives have been employed as probes to further examine the interactions between these species and Mn at the active site of H2O oxidation. Results of these studies show that the size of a hydroxylamine derivative regulates its ability to inactivate O2 evolution activity, and that this size-dependent inhibition behavior arises from the protein structure of Photosystem II. A set of anions (Cl-, F- and SO4
2-) is able to slow NH2OH and CH3NHOH inactivation of intact Photosystem II membranes by exerting a stabilizing influence on the extrinsic 23 and 17 kDa polypeptides. In contrast to this non-specific anion effect, only Cl- is capable of attenuating CH3NHOH and (CH3)2NOH inhibition in salt-washed preparations lacking the 23 and 17 kDa polypeptides. However, Cl- fails to protect against NH2OH inhibition in salt-washed membranes. These results indicate that the attack by NH2OH and its N-methyl derivatives on Mn occurs at different sites in the O2-evolving complex. The small reductant NH2OH acts at a Cl--insensitive site whereas the inhibitions by CH3NHOH and (CH3)2NOH involve a site that is Cl- sensitive. These findings are consistent with earlier studies showing that the size of primary amines controls the Cl- sensitivity of their binding to Mn in the O2-evolving complex.Abbreviation MES
4-morpholinoethanesulfonic acid
- PS II
Photosystem II 相似文献
103.
人肝刺激因子对大鼠实验性慢性肝损伤的保护作用 总被引:3,自引:0,他引:3
从健康孕妇水囊引产4─6个月龄的胎儿取肝,采用LaBrecque方法提取人肝刺激因子(hHSS)。经3H-胸腺嘧啶核苷参入肝DNA法测定其生物活性。表明此hHSS可刺激肝细胞DNA合成。采用皮下注射CCl4和饮用10%乙醇来制备慢性肝损伤动物模型,观察了hHSS的保护肝脏作用。结果表明:hHSS可使CCl4-乙醇所致慢性肝损伤大鼠的死亡率、血清谷丙转氨酶水平、肝组织中羟脯氨酸含量的升高以及肝组织中丙二醛的含量降低。肝组织切片表明:hHSS能减轻肝组织的损伤程度,促进肝细胞再生,并能明显防止肝纤维化的形成和发展。可见,hHSS对CCl4-乙醇所致的慢性肝损伤大鼠有明显的保护作用,其机制可能与促进肝细胞再生及抑制肝细胞膜的脂质过氧化有关。 相似文献
104.
用四氯化碳(CCl4)损伤正常大鼠后,采用Western印迹法和免疫组化法观察肝细胞原癌基因(c-fos/c-jun)的表达。Western印迹法表明,当成年大鼠的静息期肝细胞受到CCl4损伤性刺激后,c-fos/c-jun产物(Fos和Jun)水平升高,在CCl4处理后30min开始升高,在4h时消失。8h后Fos/Jun再度出现,并持续24h以上。ICC法表明,Jun阳性细胞为靠近肝中央静脉区的肝实质细胞。根据上述资料推测,肝受CCl4损伤后肝细胞的原癌基因c-fos/c-jun出现即时的与滞后的两次表达,这与肝细胞进入细胞周期有关,这种基因表达也许可作为肝再生过程中识别特殊体液因子的标志。 相似文献
105.
106.
单纯疱疹病毒(HSV)Ⅰ型及Ⅱ型之间有很多共同抗原,能引起血清学交叉反应,鉴别诊断比较困难。本实验利用重组DNA技术,将部分HSV-2DNA的PstI片段克隆到载体质粒PSK中,并筛选出两个重组质粒(P和P)只与HSV-2反应,与HSV-1不反应,这两个重组质粒中所含的HSV-2DNA片段大小分别是3.1和4.3kb,另外,还筛选了一个重组质粒(PHSV2-1,含5.8kbHSV-2DNA片段)与HSV-1和HSV-2均反应。将4.3kb的片段用光生物素标记后作为探针检测了159份人阴道拭子,其中23份样品呈阳性反应,其余均为阴性,从23份阳性样品中挑选12价涂片用间接荧光抗体法检测也都呈阳性反应,随机挑选的几份杂交反应阴性样品在间接荧光试验中也是阴性。本实验制备的HSV通用及HSV-2型特异性探针将比常规的血清学方法诊断和鉴别HSV-1和HSV-2感染更为可靠。 相似文献
107.
用触角电位法研究了大豆蚜、棉蚜和桃蚜对39种挥发性次生化合物的反应,结果如下:1)绿叶气味化合物及其异构体的去极化电压远强于萜烯类;具有6个碳原子的醇类和醛类是最有活性的化合物.2)大豆蚜和棉蚜对醛类和脂类的反应要比对相应的醇类强,这与桃蚜的实验结果刚好相反.3)蚜虫嗅觉系统所显示的活性与刺激化合物的结构有关,醇类和醛类刺激所引起的EAG饱和的要比不饱和的强.4)大豆春迁蚜对萜烯衍生物(醇、醛、脂)的触角电位反应高于棉蚜,而对萜烃化合物的反应刚好相反.5)各种化合物引起的EAG峰形不同;牛儿醇、癸醇和香茅醇刺激所引起的波形回复最慢,而在直链饱和醇中随着碳原子数的增加而回复速度减慢. 相似文献
108.
Molecular basis for the polymorphic forms of human serum paraoxonase/arylesterase: glutamine or arginine at position 191, for the respective A or B allozymes. 总被引:24,自引:0,他引:24
The paraoxonase/arylesterase gene is located close to the cystic fibrosis gene on chromosome 7. Human serum contains two paraoxonase/arylesterase allozymes, A and B, which differ in their substrate specificities and kinetic properties. Purified A, AB, and B esterases were digested with trypsin, and the resultant peptides were compared by high-performance liquid chromatography. The elution profiles were very similar for all three samples, except for (1) one peptide (i.e., peptide A) seen only in the A and AB profiles and (2) another peptide (i.e., peptide B) seen only in the B and AB profiles. Sequencing revealed that peptide A had glutamine at amino acid position 191, whereas peptide B was generated by cleavage on the carboxy side of position 191, presumably because there was a basic (trypsin-specific) amino acid at that position. Working independently, our laboratory and one other laboratory have sequenced the coding region for paraoxonase from human liver cDNA libraries and have identified two polymorphic sites: Arg/Gln at position 191 and Leu/Met at position 54. Using PCR amplification and direct sequencing of nucleotides in both polymorphic regions with genomic DNA, we have estimated the allelic frequencies and have determined their concordance with the serum paraoxonase allozyme phenotypes in 27 unrelated adults and in 16 members of a three-generation pedigree. Among unrelated individuals, the Met/Leu polymorphism at position 54 did not correlate with the serum esterase phenotype. In contrast, the particular amino acid at position 191 correlated perfectly with serum phenotypes: A-type individuals had Gln at position 191, and B-type individuals had Arg at position 191; AB-type serum was found only with the heterozygous (Arg/Gln) combination. Pedigree analysis showed both polymorphisms to be inherited in the expected Mendelian manner and confirmed that only the 191 polymorphism showed concordance with the serum paraoxonase/arylesterase phenotypes. 相似文献
109.
本课题合成四个氮基三乙酸单酰苯胺(IDA)类及其类似物肝胆显像螯合物。(1)用家兔做了肝胆显像扫描实验,其显像效果较好。(2)~(3)为未见报道的化合物。 相似文献
110.
本文报道在我国广西隆林壮族中发现一个罕見的HbQ复合α,β地中海贫血家系。先证者女,18岁,贫血面容,肝脾肿大。化学结构分析确证本Hb变异体为HbQ Thailand[α74(EF3)Asp→His]。血红蛋白组成以及α和β珠蛋白基因分析结果表明,先证者的珠蛋白基因型为-α~Q/-α~T复合β°/β°(IVSI-1G→T/Codon17A→T);先证者父的基因型为-‘α~Q/-复合β~O/β~A(IVSI-1G→T/β~A);先证母的基因型为-α~T/αα复合β~O/β~A(Codon17A→T/β~A)。 相似文献