Comparative structural and functional analysis of Pseudomonas aeruginosa inhibitor of vertebrate lysozyme paralogs

脊椎动物溶菌酶旁系同源物铜绿假单胞菌抑制剂的比较结构和功能分析

基本信息

批准号：
10410804
负责人：
THOMAS C LEEPER
金额：
$ 13.55万
依托单位：
KENNESAW STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-26 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10410804
关键词：
Antibiotics Bacteria Binding Biochemical Pathway Biochemistry Biological Assay Biophysics Cell Wall Complement Complement Inactivators Complex Crosslinker Cystic Fibrosis Data Collection Development Drug Design ESKAPE pathogens Enzymatic Biochemistry Enzyme Inhibition Enzyme Kinetics Enzymes Family Future Genes Glycoside Hydrolases Human Hydrolase Hydrolysis Immunity In Vitro Interferometry Knock-out Lytic Mediating Methods Microbial Genetics Multi-Drug Resistance Multiple Bacterial Drug Resistance Muramidase Opportunistic Infections Organism Pathogenicity Peptidoglycan Phenotype Plasmids Property Protein Isoforms Proteins Pseudomonas aeruginosa Research Resistance Role Small Interfering RNA Specificity Structure Training Translational Repression Underrepresented Populations Universities Vertebral column Work antimicrobial antimicrobial drug comparative crosslink cystic fibrosis patients drug discovery experimental study fighting graduate student in vivo inhibitor innovation interest knock-down liquid chromatography mass spectrometry member molecular recognition mortality mutant novel novel drug class opportunistic pathogen paralogous gene pathogen pathogenic bacteria repaired structural biology undergraduate student

Antibiotics Bacteria Binding Biochemical Pathway Biochemistry Biological Assay Biophysics Cell Wall Complement Complement Inactivators Complex Crosslinker Cystic Fibrosis Data Collection Development Drug Design ESKAPE pathogens Enzymatic Biochemistry Enzyme Inhibition Enzyme Kinetics Enzymes Family Future Genes Glycoside Hydrolases Human Hydrolase Hydrolysis Immunity In Vitro Interferometry Knock-out Lytic Mediating Methods Microbial Genetics Multi-Drug Resistance Multiple Bacterial Drug Resistance Muramidase Opportunistic Infections Organism Pathogenicity Peptidoglycan Phenotype Plasmids Property Protein Isoforms Proteins Pseudomonas aeruginosa Research Resistance Role Small Interfering RNA Specificity Structure Training Translational Repression Underrepresented Populations Universities Vertebral column Work antimicrobial antimicrobial drug comparative crosslink cystic fibrosis patients drug discovery experimental study fighting graduate student in vivo inhibitor innovation interest knock-down liquid chromatography mass spectrometry member molecular recognition mortality mutant novel novel drug class opportunistic pathogen paralogous gene pathogen pathogenic bacteria repaired structural biology undergraduate student

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项目摘要

Project Summary – Comparative structural and functional analysis of Pseudomonas aeruginosa inhibitor of vertebrate lysozyme paralogs. Thomas Leeper, PI, Kennesaw State University. The proposed integrative studies will investigate a significant regulator of peptidoglycan biochemistry in the cystic fibrosis pathogen Pseudomonas aeruginosa (PA), known to display multidrug resistance (MDR). Specifically, Inhibitor of Vertebrate Lysozyme protein 2 (IVYp2) will have its structure determined by NMR (Aim 1) and compared to its ability to inhibit hydrolase enzyme kinetics (Aim 2). While IVY proteins were originally suggested to inhibit lysozyme glycoside hydrolases, subsequent work suggested that the evolutionary origin of IVY’s may have been to inhibit lytic transglycosylases (LTs). For example, IVYp2 has no effect on lysozyme activity in enzyme inhibition assays but has been shown to inhibit LTs, at least in vitro. In order to determine which LTs are recognized in vivo enzyme inhibition assays with LTs (Aim 2) will be compared to novel applications of complementation and genetically encoded crosslinking studies in IVY knockout PA strains (Aim 3). The identified cognate IVY-LT pairs will be compared to the NMR structure of IVYp2 (Aim 1) and existing complex structures for other IVYs (i.e. IVYp1) bound to lysozyme to suggest regions important for LT molecular recognition. Graduate and undergraduate students, particularly those from underrepresented groups, will significantly participate in this research to facilitate data collection while providing them important training and advancement opportunities. These innovative studies will establish the background and workflow needed for future drug discovery efforts to develop new drug classes targeting novel biochemical pathways in MDR PA.

项目摘要 - 铜绿假单胞菌抑制剂的比较结构和功能分析脊椎动物溶菌酶旁系同源物。肯尼索州立大学PI的Thomas Leeper。拟议的综合研究将调查Pepperydoglycan生物化学的重要调节剂囊性纤维化病原体铜绿假单胞菌（PA），已知显示多药电阻（MDR）。特别是，脊椎动物溶菌酶蛋白2（IVYP2）的抑制剂将确定其结构通过NMR（AIM 1），并将其抑制水解酶动力学的能力进行比较（AIM 2）。而常春藤最初建议蛋白质抑制溶菌酶糖苷水解酶，随后的工作建议常春藤的进化起源可能是抑制裂解透明糖基酶（LTS）。例如， IVYP2对酶抑制测定中的溶菌酶活性没有影响，但已被证明可以抑制LTS，至少在体外。为了确定在体内酶抑制测定中识别哪些LT的LT LTS（AIM 2）将与新颖的完成应用进行比较，并广泛编码在常春藤基因敲除PA菌株中的交联研究（AIM 3）。确定的同源Ivy-lt对将是与IVYP2的NMR结构（AIM 1）和其他IVY的现有复杂结构相比（即 IVYP1）与溶菌酶结合，建议对LT分子识别重要的区域。研究生和本科生，尤其是来自代表性不足的小组的学生，将大大参与这项研究以促进数据收集，同时为它们提供重要的培训和进步机会。这些创新的研究将确定未来所需的背景和工作流程药物发现的努力，开发针对MDR PA中新型生化途径的新药类别。