Biosynthesis of the Immunomodulatory Molecule Capsular Polysaccharide A

免疫调节分子荚膜多糖 A 的生物合成

• 批准号: 8232369
• 负责人: JERRY M TROUTMAN
• 金额: $ 32.08万
• 依托单位: UNIVERSITY OF NORTH CAROLINA CHARLOTTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-15 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8232369
• 关键词: Affect; Anabolism; Animals; Attention; Bacteria; Bacterial Polysaccharides; Bacteroides fragilis; Biochemical; Biological Assay; Capillary Electrophoresis; Cell surface; Charge; Diphosphates; Disease; Enzymes; Future; Galactose; Genes; Genetic; Glycosyltransferase Gene; Health; High Pressure Liquid Chromatography; Homologous Protein; Human; Hydro-Lyases; Immune system; In Vitro; Inflammatory Bowel Diseases; Investigation; Kinetics; Label; Laboratories; Learning; Length; Link; Mammals; Mass Spectrum Analysis; Methods; Microbe; Modeling; Monosaccharides; Multiple Sclerosis; Mutase; NMR Spectroscopy; Nucleotides; Oligosaccharides; Pathogenesis; Pathway interactions; Phase; Phosphoenolpyruvate; Play; Polymerase; Polymers; Polysaccharides; Process; Production; Prokaryotic Cells; Protein Analysis; Proteins; Pyruvate; Reaction; Role; Route; Sequence Analysis; Series; System; Techniques; Therapeutic; Transaminases; Tritium; Work; base; chemical kinetics; enzyme activity; enzyme substrate; glycosyltransferase; insight; isoprenoid; macromolecule; method development; novel; programs; research study; sugar; tool

DESCRIPTION (provided by applicant): The role of bacterial capsular polysaccharides and the pathways that produce them are integral in human health and disease playing a fundamental role in bacterial pathogenesis as well as essential symbiotic relationships with animals. A detailed understanding of the process and factors that regulate production of these macromolecules is of utmost importance to understanding how capsular polysaccharides affect healthy and diseased states. The focus of this proposal is on capsular polysaccharide A expressed on the cell surface of the symbiotic prokaryote Bacteroides Fragilis. This is an important target for detailed investigation since studies will provide tools and approaches for understanding the role of capsular polysaccharides in symbiotic microbes that may be used in enhancing human health. In addition, this pathway represents an uncharacterized biosynthetic system that is experimentally tractable and would offer insights and new methodological approaches for deriving information on the general biosynthesis of bacterial polysaccharides in pathogenic as well as symbiotic bacteria. The gene locus responsible for the biosynthesis of the tetrasaccharide repeat unit of capsular polysaccharide A has been identified. However, unambiguous biochemical analysis of the proteins produced from this gene locus has not been performed. Gene locus analysis has identified proteins homologous to four glycosyltransferases, a galactopyranose mutase, and a sugar aminotransferase, a protein responsible for pyruvation, a flippase and a polymerase. In this program we will use chemical and kinetic approaches to identify the biosynthetic route to capsular polysaccharide A, which can only be partially predicted by sequence analysis alone. In doing so, we will learn a great deal about nucleotide diphosphate sugar modifying enzymes and oligosaccharide glycosyltransferases that are currently poorly understood. PUBLIC HEALTH RELEVANCE: The role of bacterial capsular polysaccharides and the pathways that produce them are integral in human health and disease playing a fundamental role in bacterial pathogenesis as well as essential symbiotic relationships with animals. The focus of this proposal is on capsular polysaccharide A expressed on the cell surface of the symbiotic prokaryote Bacteroides Fragilis. In this program we will use chemical and kinetic approaches to elucidate the biosynthetic route to capsular polysaccharide A, enhancing the opportunity to utilize this biomolecule as a therapeutic for diseases ranging from Multiple Sclerosis to Inflammatory Bowel Disease.

描述（由申请人提供）：细菌荚膜多糖的作用和产生它们的途径在人类健康和疾病中是不可或缺的，在细菌发病机制以及与动物的基本共生关系中起着重要作用。详细了解调节这些大分子产生的过程和因素对于了解荚膜多糖如何影响健康和疾病状态至关重要。该提案的重点是在共生原核生物脆弱拟杆菌的细胞表面上表达的荚膜多糖A。这是详细调查的重要目标，因为研究将提供工具和方法，用于了解荚膜多糖在共生微生物中的作用，可用于增强人类健康。此外，这一途径代表了一个未知的生物合成系统，是实验上听话，并将提供见解和新的方法学途径，用于获得信息的一般生物合成的细菌多糖在病原菌以及共生细菌。已经鉴定了负责荚膜多糖A的四糖重复单元的生物合成的基因位点。然而，从这个基因座产生的蛋白质的明确的生化分析尚未进行。基因位点分析已经鉴定出与四种糖基转移酶同源的蛋白质，一种吡喃半乳糖苷酶和一种糖氨基转移酶，一种负责脱水的蛋白质，一种翻转酶和一种聚合酶。在这个程序中，我们将使用化学和动力学的方法来确定生物合成途径的荚膜多糖A，这只能部分预测单独的序列分析。这样做，我们将了解大量的核苷酸二磷酸糖修饰酶和寡糖糖基转移酶，目前知之甚少。 公共卫生相关性：细菌荚膜多糖的作用和产生它们的途径在人类健康和疾病中是不可或缺的，在细菌发病机制以及与动物的基本共生关系中起着根本作用。该提案的重点是在共生原核生物脆弱拟杆菌的细胞表面上表达的荚膜多糖A。在该计划中，我们将使用化学和动力学方法来阐明荚膜多糖A的生物合成途径，提高利用这种生物分子作为治疗从多发性硬化到炎症性肠病等疾病的机会。

项目成果

Complete Tetrasaccharide Repeat Unit Biosynthesis of the Immunomodulatory Bacteroides fragilis Capsular Polysaccharide A.

• DOI: 10.1021/acschembio.6b00931
• 发表时间: 2017-01-20
• 期刊: ACS chemical biology
• 影响因子: 4
• 作者: Sharma S;Erickson KM;Troutman JM
• 通讯作者: Troutman JM

Tuning the production of variable length, fluorescent polyisoprenoids using surfactant-controlled enzymatic synthesis.

使用表面活性剂控制的酶合成来调节可变长度、荧光聚异戊二烯的生产。

• DOI: 10.1021/acs.biochem.5b00310
• 发表时间: 2015
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Troutman,JerryM;Erickson,KatelynM;Scott,PhillipM;Hazel,JosephM;Martinez,ChristinaD;Dodbele,Samantha
• 通讯作者: Dodbele,Samantha

Species differences in alternative substrate utilization by the antibacterial target undecaprenyl pyrophosphate synthase.

• DOI: 10.1021/bi500545g
• 发表时间: 2014-08-05
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Dodbele, Samantha;Martinez, Christina D.;Troutman, Jerry M.
• 通讯作者: Troutman, Jerry M.

Melanopsin elevates locomotor activity during the wake state of the diurnal zebrafish.

黑视蛋白可提高昼间斑马鱼清醒状态下的运动活动。

• DOI: 10.15252/embr.202051528
• 发表时间: 2022-05-04
• 期刊: EMBO reports
• 影响因子: 7.7