Structure of the hirulog 3-thrombin complex and nature of the S' subsites of substrates and inhibitors.

The X-ray crystallographic structure of the human alpha-thrombin complex with hirulog 3 (a potent, noncleavable hirudin-based peptide of the "hirulog" class containing a beta-homoarginine at the scissile bond), which is isomorphous with that of the hirugen-thrombin crystal structure, was solved at 2.3-A resolution by starting with a model for thrombin derived from the hirugen-thrombin complex and was refined by restrained least squares methods (R = 0.132). Residues of hirulog 3 were well-defined in the electron density, which included most of the pentaglycine linker and the C-terminal helical turn that was disordered in a related structure of thrombin with hirulog 1. The interactions of D-Phe1'-Pro2'-beta-homoArg3' with the active site of thrombin were essentially identical to those of related structures of PPACK- (D-Phe-Pro-Arg chloromethyl ketone) and hirulog 1-thrombin, with the guanidinium function of the arginyl P1 residue forming a hydrogen-bonding ion pair with Asp189 of the S1 site. A noticeable shift in the CA atom of beta-homoArg3' due to the methylene insertion displaces the scissile bond from attack by Ser195, thus imparting proteolytic stability to the beta-homoArg hirulog derivative. Resolution of the pentaglycine spacer, linking N- and C-terminal functional domains into a single oligopeptide bivalent inhibitor, permitted delineation of corresponding S' subsites of thrombin. The position of Gly4' (P1') is stabilized by three hydrogen bonds with His57, Lys60F, and Ser195, while the conformational angles maintained in a strained, nonallowed configuration for non-glycyl amino acids.(ABSTRACT TRUNCATED AT 250 WORDS)     

人体基因组随机单拷贝顺序的分离及其在染色体上的定位

本文从构建杂种细胞14-7-1的基因组文库出发,用种特异的探针分离出含有人体基因组顺序的重组子,并进一步分析了其中13个克隆,得到8个单拷贝顺序。通过与已建立的杂种细胞克隆分布板杂交以及染色体的原位杂交方法,将1个单拷贝顺序FD11-1定位在11p11-q11上。由于已经报道在11号染色体上具有3个连锁群,它们分别位于11p15、11p13和11q13上,因此,FD11-1有可能为11号染色体连锁基因图的建立提供1个有意义的座位。     

光敏核不育水稻61kD特异性蛋白质的纯化和N—端序列分析

用制备型聚丙烯酰胺凝胶电泳和制备型等电聚焦纯化了曾报道的光敏核不育水稻 (Oryza sativa)农垦 58S叶绿体的特异性蛋白质 P2 ,得到 SDS- PAGE和等电聚焦 (IEF )纯的 P2。经 SDS- PAGE和 IEF测定 ,该纯蛋白质的分子量是 61 k D,等电点是 5.8。现称 P2为 P61。氨基酸序列分析表明 P61的 N-端氨基酸序列与水稻和大麦叶绿体 ATPaseβ亚基的 N-端氨基酸序列同源。     

中国典型草原放牧草场蝗虫为害损失及经济阈值的估算(英语)

传统上估算蝗虫在放牧草场为害损失的方法几乎都是用来测定对牧草秋季产量的影响,而实际上,在估算放牧草场蝗虫为害损失及经济阈值时,牧草的现存量而非秋季产量是更应考虑的因素.本文提出了一种适合测定蝗虫对牧草现存生物量影响的新方法,即野外挂笼饲养与蝗虫种群动态相结合的估算方法,并在此基础上组建了放牧草场蝗虫种群经济阈值模型;α_0 α_1M_1_α_2M_2_ α_3S_1 α_4S_2=C其中,M_1:狭翅雏蝗生物量;M_2:宽须蚁蝗生物量;S_1:狭翅雏蝗平均个体重量,S_2:宽须蚁蝗平均个体重量;α_0-α_4:常数.同时引入蝗虫种群数量和生物量两项参数来表达蝗虫种群的发生程度.     

野生紫斑牡丹和四川牡丹种子萌发特性及与其致濒的关系

野生牡丹种子一般较小,而且不同产地之间形状、大小差异较大。其萌发特性与栽培牡丹相比也有较大差异：萌发期长达半年以上,且萌发温度在１０～１５℃为宜,超过２０℃则明显不利于生根及上胚轴生长。四个不同分布地的四川牡丹种子萌发率较一致,均在６０～７７％之间。三个不同产地的紫斑牡丹种子萌发率则相差甚远,分布于甘肃文县的萌发率达７６％,而分布于陕西略阳和湖北神农架的萌发率则分别只有１２％和４．４％,出苗率则更低。本文认为紫斑牡丹的种子特性是其在自然界处于濒危状态的重要原因之一。     

杉木、台湾杉、香杉逐步判别分析

通过对杉木、台湾杉、香杉２２个种源的叶长、叶宽、叶厚、维管束的宽与厚、栅栏组织厚、机械组织厚、表皮厚和气孔数等９个叶性状进行逐步判别分析,选取栅栏组织厚（Ｘ＿６）、叶长（Ｘ＿１）和叶厚（Ｘ＿３）三个重要因子,建立优化判别方程：Ｆ＿１（台湾杉）＝－２３５．８７１４＋１７．３１０４Ｘ＿６＋２６．３５８７Ｘ＿１＋５．９０４６Ｘ＿３Ｆ＿２（杉木）＝－５６４．９２５９＋７８．４２５４Ｘ＿６＋９３．１９２３Ｘ＿１＋０．６２９５Ｘ＿３Ｆ＿３（香杉）＝－３４９．４２０５＋６４．１９２６Ｘ＿６＋６２．６４７９Ｘ＿１＋０．８９６９Ｘ＿３定量地区分这三个树种,旨在对逐步判别分析法在树种分类上的应用进行探讨。     

Lck-dependent tyrosyl phosphorylation of the phosphotyrosine phosphatase SH-PTP1 in murine T cells.

The phosphorylation and dephosphorylation of proteins on tyrosyl residues are key regulatory mechanisms in T-cell signal transduction and are controlled by the opposing activities of protein tyrosine kinases and phosphotyrosyl phosphatases (PTPs). In T cells, several nontransmembrane protein tyrosine kinases are associated with receptors; for example, Lck is bound to the coreceptors CD4 and CD8 and becomes activated upon their stimulation. In comparison, little is known about the role of nontransmembrane PTPs in early T-cell signaling. SH-PTP1 (PTP1C, HCP, SHP) is a nontransmembrane PTP expressed primarily in hematopoietic cells, including T cells. We have found that SH-PTP1 is basally phosphorylated on serine in resting T cells. Upon stimulation of CD4 or CD8 either in a T-cell hybridoma cell line or in primary thymocytes, SH-PTP1 becomes tyrosyl phosphorylated. Moreover, SH-PTP1 is constitutively phosphorylated on tyrosine in the Lck-overexpressing lymphoma cell line LSTRA. SH-PTP1 is also a good substrate for recombinant Lck in vitro. Comparisons of the tryptic phosphopeptide maps of wild-type SH-PTP1 and deletion and point mutations establish that the two sites (Y-536 and Y-564) which are directly phosphorylated by Lck in vitro are also phosphorylated in vivo in LSTRA cells. One of these sites (Y-564) is phosphorylated in T cells in response to Lck activation. We conclude that SH-PTP1 undergoes Lck-dependent tyrosyl phosphorylation in T cells and likely plays a role in early T-cell signaling.