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951.
重组水蛭素HV2的稳定性 总被引:3,自引:0,他引:3
重组水蛭素HV2是凝血酶的特异性抑制剂,是一种非常稳定的蛋白质。温度的升高(100℃水浴)和pH(1─13)的改变不影响其活力,在某些变性剂(8mol/L尿素、1%SDS和6mol/L盐酸胍)存在的条件下也非常稳定,0.1mol/L的DTT在70℃时使其部分失活,只有pH和温度同时升高其活力才开始下降,pH13、80℃处理15min即完全失活,氨基酸组成和活性分析发现失活样品的Cys和Lys被破坏。重组水蛭素HV2含有一个结构紧密的N端核心区和一个无序的C端尾部。其N端的3个Lys-Xaa键均不被胰蛋白酶水解;胃蛋白酶及糜蛋白酶消化后,分离所得片段,氨基酸组成分析发现N端核心区依然保持很高的抗凝血酶活性,继续消化24h,核心区不被进一步降解。 相似文献
952.
汉防己甲素(汉甲)及克矽平(Polyvinylpyridine-N-Oxide,PVNO)是目前较为有效的抑制矽肺纤维化的药物。本文研究了其对胶原mRNA水平的影响.斑点杂交实验表明大鼠接尘60天和120天后α1(Ⅰ)及α1(Ⅲ)mRNA水平明显上升,经汉甲或克矽平治疗1个月或3个月后,胶原mRNA水平明显下降。原位杂交结果表明胶原mR-NA银颗粒与细胞性结节和增厚的肺泡壁的成纤维细胞分布重合。汉甲或克矽平治疗后银颗粒数下降。提示汉甲及克矽平对矽肺进程中的胶原基因表达增强有抑制作用。 相似文献
953.
954.
用奎吖咽(quinacrine)作荧光指标剂,测定玉米(ZeamaysL.)根尖微粒体(MIC)膜囊泡的H~+-泵活性,结果表明1mmol/LNaN_3仅抑制该泵活性约8%,而0.8mmol/L钒酸盐(Van)则可抑制其活性达80%,说明MIC制剂中H~+-泵活性主要由质膜(PM)H~+-ATPase产生。此泵活性严格需要Mg~(2+),二价阳离子作用大小的顺序为Mg~(2+)>Mn~(2+)>Zn~(2+)>Ca~(2+)=0;阴离子作用大小的1顺序为Br~->Cl~->NO_3~->SO_4~(2-),并初步证实当质膜同侧发生电子传递时,没有跨膜H~+梯度(△μH~+)生成。 相似文献
955.
将编码人单核细胞趋化蛋白-1(MCP-1)的基因亚克隆到大肠杆菌表达载体pEX31A中,在大肠杆菌中表达出MS2/MCP-1融合蛋0白,该表达产物约占菌体总蛋白的15%左右,Westernblot检测表明,表达产物可与MCP-1抗体特异反应。采用琼脂糖平板法进行活性测定表明,表达产物具有明显的单核细胞趋化活性,说明N端融合一段细菌蛋白对MCP-1有无趋化活性可能没有影响。 相似文献
956.
浒苔多糖剂量150mg/kg可使高胆固醇血症小鼠血清胆固醇下降22%,剂量168mg/kg可使高脂血症大鼠TCH和TG分别降低58%和61%,HDL升高27%,剂量250mg/kg可分别提高血清、脑和肝SOD活力33%、118%和224%,剂量168mg/kg对高血脂大鼠血清和心脏LPO含量降低35%和46%。 相似文献
957.
Construction of a novel bifunctional biogenic amine receptor by two point mutations of the H2-histamine receptor.
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J. DelValle I. Gantz L. Wang Y. J. Guo G. Munzert T. Tashiro Y. Konda T. Yamada 《Molecular medicine (Cambridge, Mass.)》1995,1(3):280-286
BACKGROUND: H2-histamine receptors mediate a wide range of physiological functions extending from stimulation of gastric acid secretion to induction of human promyelocyte differentiation. We have previously cloned the H2-histamine receptor gene and noted that only three amino acids on the receptor were sufficient to define its specificity and selectivity. Despite only modest overall amino acid homology (34% amino acid identity and 57.5% similarity) between the H2-histamine receptor and the receptor for another monoamine, the beta 2-adrenergic receptor, there is remarkable similarity at their critical ligand binding sites. We hypothesized that, if the specificity and selectivity of both receptors are invested in just three amino acids, it should be possible to convert one of the receptors into one that recognizes the ligand of the other by simple mutations at only one or two sites. MATERIAL AND METHODS: We explored the effect of two single mutations in the fifth transmembrane domain of the H2-histamine receptor, which encompasses the sites that determine H2 selectivity. The canine H2 receptor gene was mutated at Asp186 and Gly187 (Asp186 to Ala186 and Gly187 to Ser187) by oligonuceotide directed mutagenesis. The coding region of both the wild-type and mutated H2 receptors was subcloned into the eukaryotic expression vector, CMVneo, and stably transfected into Hepa cells and L cells. The biological activity of histamine and epinephrine on the expressed receptor was examined by measurement of cellular cAMP production and inositol trisphosphate formation. RESULTS: Hepa cells transfected with the Ala186-Ser187 mutant H2 receptor demonstrated a biphasic rise in cAMP in response to epinephrine with an early phase (ED50 approximately 10(-11) M) that could be inhibited by both propranolol and cimetidine. Epinephrine also induced IP3 generation in the same cells, a biological response that is characteristic of activation of the wild-type H2 but not of the beta-adrenergic receptor. L cells transfected with the Ala186-Ser187 mutant H2 receptor also responded to epinephrine in a cimetidine and propranolol inhibitable manner. CONCLUSIONS: We converted the H2-histamine receptor into a bifunctional one that has characteristics of both histamine and adrenergic receptors by two simple mutations. These results support the hypothesis that ligand specificity is determined by only a few key points on a receptor regardless of the structure of the remainder of the molecule. Our studies have important implications on the design of pharmacological agents targeted for action at physiological receptors. 相似文献
958.
Kan Wang Paul Drayton Bronwyn Frame Jim Dunwell John Thompson 《In vitro cellular & developmental biology. Plant》1995,31(2):101-104
A number of different methods, involving direct DNA delivery are now available for plant transformation. Here we review the most recently developed technique which involves the mixing of silicon carbide whiskers with plant cells and plasmid DNA. Fertile transgenic plants have now been produced using whisker-mediated transformation, and this method can now be considered as a simple, inexpensive alternative for plant transformation. A brief review on transformation of animal cells andChlamydomonas using whiskers technology is also included. 相似文献
959.
An RNA pseudoknot is an essential structural element of the internal ribosome entry site located within the hepatitis C virus 5' noncoding region. 总被引:8,自引:2,他引:6
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Translation of the human hepatitis C virus (HCV) RNA genome occurs by a mechanism known as "internal ribosome entry." This unusual strategy of translation is employed by naturally uncapped picornaviral genomic RNAs and several cellular mRNAs. A common feature of these RNAs is a relatively long 5' noncoding region (NCR) that folds into a complex secondary structure harboring an internal ribosome entry site (IRES). Evidence derived from the use of dicistronic expression systems, combined with an extensive mutational analysis, demonstrated the presence of an IRES within the HCV 5'NCR. The results of our continued mutational analysis to map the critical structural elements of the HCV IRES has led to the identification of a pseudoknot structure upstream of the initiator AUG. The evidence presented in this study is based upon the mutational analysis of the putative pseudoknot structure. This is further substantiated by biochemical and enzymatic probing of the wild-type and mutant 5'NCR. Further, the thermodynamic calculations, based upon a modified RNAKNOT program, are consistent with the presence of a pseudoknot structure located upstream of the initiator AUG. Maintenance of this structural element is critical for internal initiation of translation. The pseudoknot structure in the 5'NCR represents a highly conserved feature of all HCV subtypes and members of the pestivirus family, including hog cholera virus and bovine viral diarrhea virus. 相似文献
960.
家鸡和原鸡的线粒体DNA多态性比较 总被引:17,自引:0,他引:17
本文运用11种限制性内切酶分析了家鸡(茶花鸡、尼西鸡、大理漾濞黄鸡)和原鸡共10只个体的线粒体DNA限制性片段长度多态性(RFLP),平均每个个检测到的片段为40条左右。但仅发现3种变异的限制性片制性格局,即StuI-B,Eca-I-b和RI-B.其中StuI-B和ScaI-B为首次报道,而且均为原鸡所物有,EcoRI-B则为大理漾濞黄鸡所特有。茶花鸡和尼西鸡拥有完全相同的限制性格局。经过计算,原 相似文献