Assembly and export of mycobacterial lipoglycans

分枝杆菌脂聚糖的组装和输出

• 批准号: 10620764
• 负责人: Mary Jackson
• 金额: $ 60.09万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-10 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10620764
• 关键词: Abbreviations; Actinobacteria class; Acylation; Acyltransferase; Anabolism; Bacillus; Biochemical; Biochemistry; Biogenesis; Biological; Biological Assay; Blood capillaries; Cell membrane; Cell model; Cell surface; Cells; Chemicals; Complex; Corynebacterium glutamicum; Dedications; Development; Diglycerides; Disease; Dissection; Electrophoresis; Enzymes; Esterification; Event; Face; Family; Fatty Acids; Genetic; Genomics; Genus Mycobacterium; Glycerol; Glycolipids; Goals; HIV/TB; Human; Hydroxyl Radical; Immune response; Inositol; Intervention; Investigation; Knowledge; Laboratories; Leprosy; Ligands; Lipopolysaccharides; Location; Mannans; Mannose; Mannosides; Mannosyltransferases; Mass Spectrum Analysis; Membrane; Metabolic; Molecular; Multi-Drug Resistance; Mycobacterium tuberculosis; Nature; Nomenclature; Open Reading Frames; Organism; PIM1 gene; Pathogenesis; Pathway interactions; Phagocytes; Pharmaceutical Preparations; Phenotype; Phosphatidylinositols; Physiology; Play; Prevalence; Prevention; Process; Recombinants; Research; Resort; Role; Side; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Streptomyces coelicolor; Structure; Structure-Activity Relationship; Therapeutic Intervention; Thin Layer Chromatography; Time; Tuberculosis; Vaccines; Vertebral column; Virulence Factors; Xylose; acyl group; arabinogalactan; cell envelope; co-infection; deoxycholate; design; drug development; enzyme activity; enzyme pathway; epimerase; extensive drug resistance; genetic manipulation; glycosylation; glycosyltransferase; immunoregulation; inhaled nitric oxide; interest; lipoarabinomannan; lipomannan; mutant; mycobacterial; myoinositol; novel; novel therapeutics; pathogen; phosphatidylinositol mannoside; prophylactic; targeted treatment; treatment strategy; tuberculosis drugs

Abstract Phosphatidylinositol mannosides (PIM) and their multiglycosylated counterparts, lipomannan (LM) and lipoarabinomannan (LAM), are complex glycolipids and lipoglycans found in the cell envelopes of all mycobacterial species. They play various essential although poorly defined roles in mycobacterial physiology and are important immunomodulatory molecules in the course of tuberculosis and leprosy as well as key ligands promoting the entry of mycobacteria and their survival within phagocytic and non-phagocytic cells. Although much progress has been made over the last 25 years in elucidating the structures and biosynthesis of these molecules, fundamental questions remain about the pathways leading to their biosynthesis and translocation to the cell surface. Furthermore, while the pleiotropic biological activities displayed by purified PIM, LM and LAM in cellular models suggest that they play important roles in pathogenesis, studies aimed at validating this assumption and precisely delineating their contribution to host-pathogen interactions when carried by intact bacilli are still limited by the paucity of mutants deficient in well-defined aspects of the biosynthesis and export of these molecules that are available. We propose to pursue structural, genetic and biochemical studies toward a complete definition of the structure (Aim 1), biosynthesis (Aim 2) and export (Aim 3) of PIM, LM and LAM. Completing our understanding of PIM, LM and LAM biogenesis, in addition to providing fundamental knowledge about the biochemistry of Mycobacterium tuberculosis (Mtb), is expected to lead to the discovery of essential enzymes and transporters which, much like the arabinosyltransferases of the Emb family and the epimerase DprE1, could provide new opportunities for anti-tuberculosis drug development. The availability of recombinant strains accumulating structurally defined biosynthetic precursors will facilitate structure-function relationship studies, and that of defined Mtb mutants deficient in various aspects of PIM, LM and LAM synthesis will allow a precise assessment of the contribution of these molecules to the immunopathogenesis of tuberculosis. Abbreviations: AG, arabinogalactan; AM, arabinomannan; AcylT, acyltransferase; Araf, arabinofuranosyl; AraT, arabinosyltransferase; CZE, capillary zone electrophoresis; DOC, deoxycholate; GT, glycosyltransferase; Ino, myo-Inositol; LAM, lipoarabinomannan; LM, lipomannan; LPS, lipopolysaccharide; MALDI-TOF, Matrix-Assisted Laser desorption/ionization time of flight; Manp, mannopyranosyl; ManT, mannosyltransferase; MPI, mannosylated phosphatidylinositol; MS, mass spectrometry; MTX, methyl-thio-xylose; ORF, open reading frame; OM, outer membrane; PIM, phosphatidyl-myo-inositol mannosides; TLC, thin-layer chromatography. Nomenclature: PIM is used to describe the global family of phosphatidylinositol mannosides that carries one to four fatty acids (attached to the glycerol, inositol and/or mannose) and one to six mannose residues. In AcXPIMY, x refers to the number of acyl groups esterified to available hydroxyls on the mannose or myo-inositol residues, y refers to the number of mannose residues; e.g. Ac1PIM1 corresponds to the phosphatidylinositol mono-mannoside PIM1 carrying two acyl groups attached to the glycerol (the diacylglycerol substituent) and one acyl group esterified to the mannose residue.

抽象的 磷脂酰肌醇甘露糖苷（PIM）及其多糖基化对应物，Lipomannan（LM）和 脂肪芳替兰南南（LAM）是在所有细胞信封中发现的复杂的糖脂和脂肪糖果 分枝杆菌。他们在分枝杆菌生理学中发挥各种必不可少的作用 并且是结核病和麻风病以及钥匙的重要免疫调节分子 促进分枝杆菌进入的配体及其在吞噬细胞和非吞噬细胞中的生存。 尽管在过去的25年中取得了很大的进步，以阐明结构和生物合成 在这些分子中，关于导致其生物合成的途径和 转移到细胞表面。此外，纯化的多效性生物活性 细胞模型中的PIM，LM和LAM表明它们在发病机理中起着重要作用，针对 验证这一假设并精确地描绘出其对宿主病原体相互作用的贡献 由完整的杆菌携带的仍然受到突变体在明确明确的方面缺乏的限制 可用的这些分子的生物合成和输出。 我们建议从结构，遗传和生化研究进行完整的结构定义 （AIM 1），PIM，LM和LAM的生物合成（AIM 2）和出口（AIM 3）。完成我们对PIM的理解， LM和LAM生物发生，除了提供有关生物化学的基本知识 结核分枝杆菌（MTB）有望导致发现必需酶和转运蛋白 就像EMB家族的阿拉伯糖基转移酶和分配酶DPRE1一样，可以提供新的 抗结核药物开发的机会。重组菌株的可用性积累 结构定义的生物合成前体将促进结构功能关系研究，以及 定义的MTB突变体在PIM，LM和LAM合成的各个方面缺陷，将允许精确 评估这些分子对结核病的免疫发病发生的贡献。 缩写： AG，Arabinogalactan； Am，Arabinomannan；酰基，酰基转移酶； Araf，阿拉伯呋喃糖基； ARAT，阿拉伯糖基转移酶； cze，毛细管 区电泳； DOC，脱氧胆酸； GT，糖基转移酶； Ino，肌醇；林，脂肪动物群； LM，Lipomannan； LPS， 脂多糖； MALDI-TOF，基质辅助激光解吸/电离时间； manp，mannopyranosyl;曼特， 甘露糖基转移酶； MPI，甘露糖基化的磷脂酰肌醇； MS，质谱； MTX，甲基 - 硫代二甲糖； ORF，开放阅读 框架; OM，外膜； PIM，磷脂酰 - 甲状腺醇甘露糖苷； TLC，薄层色谱。 命名法： PIM用于描述载有1-4种脂肪酸的全球磷脂酰肌醇甘露糖苷（附着在甘油， 肌醇和/或甘露糖）和一到六个甘露糖残留物。在acxpimy中，x是指对可用的酰基酯酯的数量 甘露糖或肌醇残基上的羟基是指甘露糖残基的数量。例如AC1PIM1对应于 磷脂酰肌醇单甘露糖苷PIM1携带两个附着在甘油（二酰基甘油取代基）和一个 酰基酯化为甘露糖残基。

项目成果

Acylation of glycerolipids in mycobacteria.

• DOI: 10.1038/s41467-023-42478-x
• 发表时间: 2023-10-23
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Angala SK;Carreras-Gonzalez A;Huc-Claustre E;Anso I;Kaur D;Jones V;Palčeková Z;Belardinelli JM;de Sousa-d'Auria C;Shi L;Slama N;Houssin C;Quémard A;McNeil M;Guerin ME;Jackson M
• 通讯作者: Jackson M

Mechanical morphotype switching as an adaptive response in mycobacteria.

• DOI: 10.1126/sciadv.adh7957
• 发表时间: 2024-01-05
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Eskandarian, Haig Alexander;Chen, Yu-Xiang;Toniolo, Chiara;Belardinelli, Juan M.;Palcekova, Zuzana;Hom, Lesley;Ashby, Paul D.;Fantner, Georg E.;Jackson, Mary;Mckinney, John D.;Javid, Babak
• 通讯作者: Javid, Babak