Characterization of a novel AB5 cytotoxin

新型 AB5 细胞毒素的表征

• 批准号: 7996641
• 负责人: Adrienne W Paton
• 金额: $ 25.7万
• 依托单位: UNIVERSITY OF ADELAIDE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-15 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7996641
• 关键词: Affect; Animal Model; Antibodies; Apoptosis; Bacteria; Bacterial Toxins; Binding; Biology; Bioterrorism; Blood; Blood Cells; Categories; Cell Death; Cell Line; Cell surface; Cells; Cellular biology; Child; Cholera Toxin; Collaborations; Computer Simulation; Confocal Microscopy; Coupled; Crystallization; Crystallography; Cultured Cells; Cytosol; Cytotoxin; Data; Development; Disease; Disease Outbreaks; Dose; Electron Microscopy; Emerging Communicable Diseases; Endoplasmic Reticulum; Escherichia coli; Eukaryotic Cell; Family; Fluorescence Microscopy; Foundations; Funding; Future; Genes; Glycolipids; Goals; Harvest; Hemolytic-Uremic Syndrome; Histologic; Histopathology; Housing; Human; Immunohistochemistry; In Vitro; Institution; Investigation; Knock-out; Knowledge; Label; Lead; Life; Microscopy; Modeling; Molecular; Molecular Chaperones; Mus; Mutate; Names; Nature; Neurologic Manifestations; Oral; Organ; Outcome; Pathogenesis; Pathology; Pathway interactions; Pertussis Toxin; Pharmaceutical Preparations; Plasmids; Positioning Attribute; Proteins; Proteomics; Publishing; Reagent; Recombinants; Reporting; Research; Research Personnel; Resistance; Resolution; Role; Serine Protease; Shiga Toxin; Specificity; Stress; Structure; Targeted Toxins; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Toxic effect; Toxin; Vero Cells; Virulence Factors; Western Blotting; X-Ray Crystallography; base; biodefense; cell injury; cellular targeting; cytotoxic; cytotoxicity; design; gastrointestinal; gastrointestinal infection; gene cloning; glycolipid receptor; holotoxins; human disease; in vivo; insight; laboratory facility; mouse model; mutant; novel; pathogen; programs; receptor; response; retrograde transport; small molecule; therapeutic vaccine; three dimensional structure; tissue tropism; tool; trafficking; uptake

We have recently discovered a novel "Subtilase cytotoxin" produced by a strain of Shiga toxigenic Escherichia coli (a category B Priority Pathogen) that was responsible for an outbreak of hemolytic uremic syndrome (HUS). It belongs to a new class of AB5 cytotoxins, because its A subunit has distinct enzymic activity and it has no sequence similarity with any of the other AB5 toxin families (cholera, Shiga and pertussis toxins). Subtilase cytotoxin is extraordinarily cytotoxic for cultured cells and is lethal for mice (with histopathology similar to that of HUS). Apart from its possible role in severe human disease, it is a potential bioterrorism agent which could have global impact. The long-term objective of this response to PA-04-119 is complete structural and functional characterization of the emerging toxin. Its Specific Aims are: 1. Elucidation of the molecular basis for cytotoxicity. A subunit Subtilase-like serine protease activity is essential for cytotoxicity, and preliminary data indicate that the ER chaperone BiP is a key cellular target for the toxin. In this Aim, we will continue our investigation of the molecular basis for cytotoxicity by examining the downstream consequences of BiP cleavage, including whether this results in ER stress and apoptosis. 2. Investigation of toxin trafficking in target cells. We hypothesize that after binding to the cell surface, the toxin must undergo retrograde transport to the ER in order to exert its toxic effects. Intracellular toxin trafficking will therefore be examined by co-localization of labelled toxin with cellular compartmental markers using confocal microscopy. 3. Examination of tissue tropism and in vivo effects of the toxin. The distribution of toxin receptors in mouse tissues and trafficking of labelled toxin in vivo will be examined using fluorescence, confocal and electron microscopy. The direct contribution of Subtilase cytotoxin to the pathogenesis of STEC disease will also be investigated in a mouse model. 4. Determination of the 3Dstructure of the toxin. X-ray crystallography and computer modelling will be used to solve the 3D structure. Relevance: This project will provide essential information on the basic biology of this emerging toxin, particularly its structure and the mechanism whereby it damages cells and tissues. This will provide a foundation for the future development of countermeasures including therapeutic agents and vaccines.

我们最近发现了一种新的“枯草酶细胞毒素”，它是由一种志贺毒素菌株产生的

项目成果

Incorporation of a non-human glycan mediates human susceptibility to a bacterial toxin.

• DOI: 10.1038/nature07428
• 发表时间: 2008-12-04
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Byres, Emma;Paton, Adrienne W.;Paton, James C.;Lofling, Jonas C.;Smith, David F.;Wilce, Matthew C. J.;Talbot, Ursula M.;Chong, Damien C.;Yu, Hai;Huang, Shengshu;Chen, Xi;Varki, Nissi M.;Varki, Ajit;Rossjohn, Jamie;Beddoe, Travis
• 通讯作者: Beddoe, Travis

Structure, biological functions and applications of the AB5 toxins.

• DOI: 10.1016/j.tibs.2010.02.003
• 发表时间: 2010-07
• 期刊: TRENDS IN BIOCHEMICAL SCIENCES
• 影响因子: 13.8
• 作者: Beddoe, Travis;Paton, Adrienne W.;Le Nours, Jerome;Rossjohn, Jamie;Paton, James C.
• 通讯作者: Paton, James C.

Escherichia coli Subtilase Cytotoxin.

• DOI: 10.3390/toxins2020215
• 发表时间: 2010-02-01
• 期刊: Toxins
• 影响因子: 4.2
• 作者: Paton AW;Paton JC
• 通讯作者: Paton JC

Activation of the Akt-NF-kappaB pathway by subtilase cytotoxin through the ATF6 branch of the unfolded protein response.

• DOI: 10.4049/jimmunol.0900017
• 发表时间: 2009-07-15
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Yamazaki H;Hiramatsu N;Hayakawa K;Tagawa Y;Okamura M;Ogata R;Huang T;Nakajima S;Yao J;Paton AW;Paton JC;Kitamura M
• 通讯作者: Kitamura M

Tissue factor–dependent procoagulant activity of subtilase cytotoxin, a potent AB5 toxin produced by shiga toxigenic Escherichia coli.

枯草杆菌酶细胞毒素的组织因子依赖性促凝血活性，枯草杆菌酶细胞毒素是由产志贺毒素大肠杆菌产生的强效 AB5 毒素。

• DOI: 10.1086/656534
• 发表时间: 2010
• 期刊: The Journal of infectious diseases
• 作者: Wang,Hui;Paton,JamesC;Thorpe,ChelesteM;Bonder,ClaudineS;Sun,WaiYan;Paton,AdrienneW
• 通讯作者: Paton,AdrienneW