Karyotypic evolution of the family Sciuridae: inferences from the genome organizations of ground squirrels

Cross- species chromosome painting has made a great contribution to our understanding of the evolution of karyotypes and genome organizations of mammals. Several recent papers of comparative painting between tree and flying squirrels have shed some light

A NOVEL PLASMINOGEN-ACTIVATOR FROM SNAKE-VENOM - PURIFICATION, CHARACTERIZATION, AND MOLECULAR-CLONING

A novel plasminogen activator from Trimeresurus stejnegeri venom (TSV-PA) has been identified and purified to homogeneity. It is a single chain glycoprotein with an apparent molecular weight of 33,000 and an isoelectric point of pH 5.2. It specifically activates plasminogen through an enzymatic reaction. The activation of human native GIu-plasminogen by TSV-PA is due to a single cleavage of the molecule at the peptide bond Arg(561)-Val-(562). Purified TSV-PA, which catalyzes the hydrolysis of several tripeptide p-nitroanilide substrates, does not activate nor degrade prothrombin, factor X, or protein C and does not clot fibrinogen nor show fibrino(geno)lytic activity in the absence of plasminogen. The activity of TSV-PA was readily inhibited by phenylmethanesulfonyl fluoride and by p-nitrophenyl-p-guanidinobenzoate. Oligonucleotide primers designed on the basis of the N-terminal and the internal peptide sequences of TSV-PA were used for the amplification of cDNA fragments by polymerase chain reaction. This allowed the cloning of a full-length cDNA encoding TSV-PA from a cDNA library prepared from the venom glands. The deduced complete amino acid sequence of TSV-PA indicates that the mature TSV-PA protein is composed of 234 amino acids and contains a single potential N-gIycosylation site at Asn(1G1). The sequence of TSV-PA exhibits a high degree of sequence identity with other snake venom proteases: 66% with the protein C activator from Aghistrodon contortrix contortrix venom, 63% with batroxobin, and 60% with the factor V activator from Russell's viper venom. On the other hand, TSV-PA shows only 21-23% sequence similarity with the catalytic domains of u-PA and t-PA. Furthermore, TSV-PA lacks the sequence site that has been demonstrated to be responsible for the interaction of t-PA (KHRR) and u-PA (RRHR) with plasminogen activator inhibitor type 1.

Trimeresurus stejnegeri snake venom plasminogen activator - Site-directed mutagenesis and molecular modeling

The specific plasminogen activator from Trimeresurus stejnegeri venom (TSV-PA) is a serine proteinase presenting 23% sequence identity with the proteinase domain of tissue type plasminogen activator, and 63% with batroxobin, a fibrinogen clotting enzyme from Bothrops atrox venom that does not activate plasminogen. TSV-PA contains six disulfide bonds and has been successfully overexpressed in Escherichia coli (Zhang, Y., Wisner, A., Xiong, Y. L,, and Bon, C, (1995) J. Biol. Chem. 270, 10246-10255), To identify the functional domains of TSV-PA, we focused on three short peptide fragments of TSV-PA showing important sequence differences with batroxobin and other venom serine proteinases. Molecular modeling shows that these sequences are located in surface loop regions, one of which is next to the catalytic site, When these sequences were replaced in TSV-PA by the equivalent batroxobin residues none generated either fibrinogen-clotting or direct fibrinogenolytic activity, Two of the replacements had little effect in general and are not critical to the specificity of TSV-PA for plasminogen. Nevertheless, the third replacement, produced by the conversion of the sequence DDE 96a-98 to NVI, significantly increased the K-m for some tripeptide chromogenic substrates and resulted in undetectable plasminogen activation, indicating the key role that the sequence plays in substrate recognition by the enzyme.

碱基互变异构体对碱基配对影响的量子生物学研究

通过分子轨道法计算, 按能量极小化原理,就碱基互变异构体对碱基配对的影响作了量子生物学研究。结果表明, 互变异构体的存在可以使碱基间呈现多种配对方式。例如G的醇型异构体G(OH)的存在, 一方面可能导致基因自发突变,另一方面可能增加DNA双链结构的稳定性,影响转录。此外,对“氢键专一性”的说法提出了异议。图3表3参15

绵羊大肠阻塞病理模型血浆及组织中内皮素-1含量的变化

选择绵羊建立反刍动物大肠阻塞病理模型,对手术前、后静脉血浆及死亡后心脏、肝脏、脾脏、肺脏、肾脏、胰脏和肠组织中内皮素-1(ET-1)含量进行测定,探讨了该模型血浆及组织中ET-1含量的变化和临床意义。结果表明:模型组和对照组手术前血浆ET-1含量分别为(57.81±2.95)ng/L和(62.72±3.17)ng/L,手术后含量分别为(41.87±8.06)ng/L和(60.74±4.68)ng/L,模型组术后ET--1含量明显下降(P<0.05);每30min间隔采血,模型组术前、术后第2天和第4天ET--1含量依次降低,差异极显著(P<0.01);模型组心、肺、肾组织中的ET-1含量分别为(273.28±53.7)、(270.65±98.6)、(249.32±20.8)ng/L,分别比对照组心脏(105.73±32.2)ng/L、肺脏((34.34±6.73)ng/L、肾脏(50.43±12.89)ng/L明显升高(P<0.01)。这些变化可能与肠阻塞的发生、发展有密切关系。

用神经网络法预测膜蛋白RH和BR的二级结构

以视紫红质和菌紫质两个典型的膜蛋白为选材, 用神经网络方法对其二级结构进行预测。预测结果与实验资料的符合率与该方法用于球蛋白时的结果一致。由此暗示蛋白质二级结构主要由蛋白质内部 相互作用所决定, 而外部环境不起主导作用。表2参13

p53蛋白质C端的三维结构及生物功能区

利用p53 C端118个氨基酸的mRNA二级结构和Chou-Fasman蛋白质二级结构预测原则,预测p53蛋白质C端289-325为卷曲肽段,368-393段包括两段螺旋结构:#alpha#_(1)368-373、#alpha#_(2)381-388。其中三段已知的蛋白质二级结构与此mRNA二级结构单元间有准确的对应关系。与四种以多重序列联配为基础的蛋白质二级结构预测方法(准确率均为73.20%左右)相对照,预测结果基本一致。结合单体聚合区31个氨基酸晶体结构,在SGI INDIGO~(2)工作站上构建了p53 C端108个残基的三维结构。进一步揭示了p53 C端诸多生物功能区之间的空间构象关系。

The three dimensional structure and biological domains of the p53 C-terminus.

It was expected that there are a coil (289 similar to 325) and two a helix (alpha(1)368 similar to 373, alpha(2)381 similar to 388) structures in p53 protein C-terminal region based on its mRNA secondary structure template and Chou-Fasman's protein secondary structure principle of prediction. The result was conformed by the other four methods of protein secondary structure prediction that are based on the multiple sequence alignment (accuracy = 73.20%). Combine with the 31 amino acids crystal structure of the oligomerization, the three dimensional conformation of p53 C-terminal 108 residues was built using the SGI INDIGO(2) computer. This structure further expounds the relationship among those biological function domains of p53 C- terminus at three-dimensional level.

Sleeping sites of black-and-white snub-nosed monkeys (Rhinopithecus bieti) at Baima Snow Mountain, China

Data on sleeping sites of a group of black-and-white snub-nosed monkeys Rhinopithecus bieti (Colobinae, Primates) were collected between April-July and September-December 2001 to try to determine the factors affecting site selection at Nanren (99 degrees