Zinc metalloprotease families in Trypansoma brucei

布氏锥虫中的锌金属蛋白酶家族

• 批准号: 7596894
• 负责人: John E Donelson
• 金额: $ 34.3万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7596894
• 关键词: 3' Untranslated Regions; 5' Untranslated Regions; Abbreviations; Affect; African; Amino Acid Sequence; Amino Acids; Binding; Biological; Biological Process; Blood Circulation; C-terminal; Cells; Cellular Membrane; Cleaved cell; Complement; Cyclic AMP; Cytolysis; Cytoplasm; Data; Development; Enabling Factors; Endoplasmic Reticulum; Environment; Family; Fractionation; GPI Membrane Anchors; Gene Deletion; Gene Family; Genes; Genetic Translation; Glycosylphosphatidylinositols; Green Fluorescent Proteins; Growth; Helminths; Homologous Gene; Insecta; Intercistronic Region; Label; Lead; Leishmania; Leishmania glycoprotein gp63; Life Cycle Stages; Location; Mediating; Membrane Glycoproteins; Messenger RNA; Metabolic; Metabolism; Metalloproteases; Molecular; Mutagenesis; Names; Nature; Parasites; Peptide Hydrolases; Phagolysosome; Phospholipase C; Play; Property; Proteins; Proteolysis; Protozoa; RNA Interference; Recombinants; Regulation; Reporting; Resistance; Role; Staging; Surface; System; Techniques; Tetracyclines; Translations; Trypanosoma; Trypanosoma brucei brucei; Untranslated Regions; Variant; Zinc; base; beta-Galactosidase; extracellular; mRNA Stability; macrophage; overexpression; poly(glutamic acid-lysine); prevent; research study; trafficking

DESCRIPTION (provided by the applicant): Proteases are critical factors enabling many protozoa and helminths to parasitize and survive in both mammalian and insect hosts. The biological properties and differential expression of related groups of zinc metalloproteases in the African trypanosome Trypanosoma brucei will be studied using the techniques of traditional gene deletion, RNA interference (RNAi), and overexpression for selected experiments. Three groups of metalloproteases, called TbMSP-A, TbMSP-B and TbMSP-C, each has about 33% amino acid identity with the major surface protease (MSP, formerly called GP63) of Leishmania, and share a common zinc binding motif. The mRNAs of all three gene families occur in bloodstream trypanosomes, whereas only the mRNA of TbMSP-B is expressed in procyclic trypanosomes. Their C-terminal amino acid sequences suggest that TbMSP-A and -B are likely associated with a cellular membrane via a glycosylphosphatidylinositol (GPI) anchor, whereas TbMSP-C may be either located in the cytoplasm or secreted. We hypothesize that those TbMSPs predicted on the basis of their sequences to traffic to the trypanosome surface, i.e., TbMSP-A and TbMSP-B, serve important functions in facilitating survival of trypanosomes in the host environment. Our preliminary data lead us to hypothesize that TbMSP-B is a surface protease under translational control in bloodstream cells, which participates in shedding of the variant surface glycoprotein (VSG) during differentiation from bloodstream to procyclic cells. Further evidence leads us to hypothesize that TbMSP-A serves a distinct proteolytic function that is not required in procyclic cells, although the exact nature of this function has yet to be identified. In this application we propose to use the techniques of immunolocalization, sub-cellular fractionation, metabolic labeling, protease characterization, and gene mutagenesis to identify the sub-cellular locations of the TbMSP-A and TbMSP-B families, to determine their biological functions, and to elucidate the molecular mechanisms regulating their differential expression.

描述（由申请人提供）：蛋白酶是使许多原生动物和蠕虫在哺乳动物和昆虫宿主中寄生和生存的关键因素。将使用传统基因缺失，RNA干扰（RNAI）和过表达的技术研究非洲锥虫锥虫中相关锌金属蛋白酶相关组的生物学特性和差异表达。三组金属蛋白酶，称为TBMSP-A，TBMSP-B和TBMSP-C，每个金属蛋白酶具有约33％的氨基酸同一性，与Leishmania的主要表面蛋白酶（MSP，以前称为GP63），并具有共同的锌结合基序。所有三个基因家族的mRNA都出现在血液锥虫中，而只有TBMSP-B的mRNA在procyclic锥虫中表达。它们的C末端氨基酸序列表明，TBMSP-A和-B可能通过糖基磷脂酰肌醇（GPI）锚固剂与细胞膜有关，而TBMSP-C可以位于细胞质或分泌中。我们假设那些TBMSP是根据其序列序列到锥虫表面的序列进行预测的，即TBMSP-A和TBMSP-B在促进宿主环境中锥虫存活方面起着重要功能。 我们的初步数据使我们假设TBMSP-B是血流细胞转化控制下的表面蛋白酶，该蛋白酶参与了从血液和procyclic细胞区分开的变异表面糖蛋白（VSG）的脱落。进一步的证据使我们假设TBMSP-A具有独特的蛋白水解功能，尽管尚未确定该功能的确切性质，但在procyclic细胞中不需要。在此应用中，我们建议使用免疫定位，亚细胞分馏，代谢标记，蛋白酶表征和基因诱变的技术来识别TBMSP-A和TBMSP-B家族的亚细胞位置，以确定其生物学功能，以确定其生物学机械的不同表达。

项目成果

Analysis of expressed sequence tags from the four main developmental stages of Trypanosoma congolense.

• DOI: 10.1016/j.molbiopara.2009.06.004
• 发表时间: 2009-11
• 期刊: MOLECULAR AND BIOCHEMICAL PARASITOLOGY
• 影响因子: 1.5
• 作者: Helm, Jared R.;Hertz-Fowler, Christiane;Aslett, Martin;Berriman, Matthew;Sanders, Mandy;Quail, Michael A.;Soares, Marcelo B.;Bonaldo, Maria F.;Sakurai, Tatsuya;Inoue, Noboru;Donelson, John E.
• 通讯作者: Donelson, John E.