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Conformation and functional dynamics of a bacterial PASTA kinase

细菌 PASTA 激酶的构象和功能动力学

基本信息

批准号：
10526288
负责人：
CHRISTOPHER J KRISTICH
金额：
$ 33.88万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10526288
关键词：
Address Adopted Antibiotic Resistance Antibiotics Antimicrobial Resistance Bacteria Bile fluid Binding Biological Biological Adaptation Biological Assay Biology Cell Wall Cell division Cephalosporins Cytoplasm Cytoplasmic Tail Detergents Development Dimerization Disulfides Electron Spin Resonance Spectroscopy Enterococcus Enterococcus faecalis Family Foundations Goals Gram-Positive Bacteria Hospitals Impairment In Vitro Individual Infection Knowledge Length Mediating Membrane Molecular Molecular Conformation Mutagenesis Mutation NMR Spectroscopy Nature Nosocomial Infections Pathogenesis Pathway interactions Phosphorylation Phosphotransferases Positioning Attribute Prior Therapy Process Production Protein Family Public Health Regulation Reporter Research Resistance Risk Factors Role Scheme Signaling Protein Site Staphylococcus aureus Stimulus Structure Structure-Activity Relationship Therapeutic Toxin Transmembrane Domain Virulence Work X-Ray Crystallography acquired factor antimicrobial antimicrobial drug assault colonization resistance crosslink design extracellular genetic selection gut colonization gut microbiota in vivo innovation insight member mutant new therapeutic target novel novel strategies novel therapeutic intervention pathogen pathogenic bacteria response trait

项目摘要

Project Summary/Abstract Enterococci such as Enterococcus faecalis are among the trillions of intestinal microbes that normally coexist in a symbiotic relationship with their host. However, antibiotic-resistant enterococci are also among the three most common causes of hospital-acquired infections and therefore represent a serious public health problem. An important risk factor for the acquisition of hospital-acquired enterococcal infections is prior therapy with antibiotics, such as broad-spectrum cephalosporins, to which enterococci are intrinsically resistant. Thus, the inherent abilities of enterococci to colonize the gut and withstand assault by antimicrobial agents are especially critical for pathogenesis of enterococcal infections. Yet, the mechanisms of enterococcal gut colonization and antimicrobial resistance are not fully understood. One critical determinant of these intrinsic traits is a transmembrane Ser/Thr kinase known as IreK, which belongs to a family of bacterial kinases containing extracellular PASTA domains. Such “PASTA kinases” control critical processes including antibiotic resistance, toxin production, virulence, or cell division in nearly all Gram-positive bacteria; in some bacteria, a PASTA kinase is essential for viability. As such, PASTA kinases represent attractive targets for new therapeutics. However, a basic understanding of the structure and dynamics of PASTA kinases in solution, and the structure-function relationships that promote environmental sensing and coordination of biological responses in vivo, is lacking. Such information is critical to inform development of new therapeutic approaches. The research proposed here seeks to address this gap by elucidating fundamental structure-function relationships in solution for a representative PASTA kinase, the IreK kinase from E. faecalis, which we have shown is required for intrinsic resistance of E. faecalis to cell-wall-active antibiotics and to detergents present in bile. This collaborative research is designed to apply new approaches and perspectives to elucidate novel insights into the function of the IreK kinase using a complementary combination of in vivo functional assays and EPR spectroscopy approaches to determine the PASTA module conformation in solution and understand how kinase domain dynamics contribute to kinase activation. By successfully completing this work, we will obtain important new insights into a representative bacterial PASTA kinase to help define the structure-function relationships for an entire family of critical bacterial signaling proteins with the long-term goal of modulating the activity of these kinases therapeutically.

项目总结/文摘