BIOCHEMICAL CYTOLOGY OF HOST-PARASITE INTERACTIONS IN PARASITIC PROTOZOA

利什曼原虫和锥虫的细胞生物学和生物化学 作为细胞内和细胞外寄生的模型被研究， 分别进行了分析。因为宿主和寄生虫之间的所有相互作用都发生在 在寄生虫表面膜(SM)的水平上，重点是 放在：1)集生化和生理为一体 特征和2)确定其在寄生虫存活和 发展。 一种76 KDa的含羧基载体的SM糖蛋白 杜氏乳杆菌(L.donovani，LD)前鞭毛体的功能被鉴定。A SM L-含两个41亚基的脯氨酸结合/转运蛋白 在LD中鉴定出34 KDa。生物合成的分泌物 可溶性酸性磷酸酶途径(S-酸性磷酸酶)在两者中均有定位 我是支持和迷信的。S-乙酰胆碱得到提纯，前30个 测定了其N端氨基酸序列的残基。 基于该序列的寡核苷酸探针用于 鉴定并克隆S酸性磷酸酶基因。可诱导LD SM 3‘- 核苷酸酶/外切核糖核酸酶的分离、纯化和鉴定 是一种43 kDa的甘露糖化糖蛋白。抗体正在被 用于筛选表达文库以鉴定和克隆ITS 吉恩。LD SM 5‘-核苷酸酶的分离、纯化和鉴定 是一种含有两个72亚基的甘露糖化糖蛋白 和43 kDa。此外，LD SM葡萄糖转运蛋白和 对质子-ATPase进行了鉴定和性质研究。基于他们的 与其他系统中类似蛋白质的同源性， 合成了保守功能区的寡核苷酸 作为鉴定和克隆这两个基因的探针 利什曼蛋白。 目前的结果提供了进一步的功能表征 寄生的SM成分，这些成分可能是潜在的 化疗和/或免疫诊断/预防的目标。

The cell biology and biochemistry of Leishmania and Trypanosoma are investigated as models of intra- and extracellular parasitism, respectively. As all interactions between host and parasite occur at the level of the parasite surface membrane (SM), emphasis is placed on: 1) its integrated biochemical and physiological characterization and 2) defining its roles in parasite survival and development. A 76 KDa SM glycoprotein, with putative carboxyl-carrier functions was identified in L. donovani (Ld) promastigotes. A SM L-proline-binding/transport protein containing two subunits of 41 and 34 KDa was identified in Ld. The biosynthetic secretory pathway of soluble-acid phosphatase (S-AcP) was localized in both Ld pro- and amstigotes. S-AcP was purified and the first 30 residues of its N-terminal amino acid sequence determined. Oligonucleotide probes based on this sequence were used to identify and clone the S-AcP gene. The inducible Ld SM 3'- nucleotidase/exo-ribonuclease was isolated, purified and shown to be a 43 kDa mannosylated glycoprotein. Antibodies are being used to screen cDNA-expression libraries to identify and clone its gene. The LD SM 5'-nucleotidase was isolated, purified and shown to be a mannosylated glycoprotein containing two subunits of 72 and 43 kDa. Further, the LD SM glucose transport protein and proton-ATPase were identified and characterized. Based on their homologies with similar proteins in other systems, oligonucleotides of conserved functional regions were synthesized as probes for identifying and cloning the genes for these two leishmanial proteins. The current results provide further functional characterization of parasite SM constituents and these could serve as potential targets for chemotherapy and/or immunodiagnosis/-prophylaxis.