Structural characterization of MCE mediated lipid transport across the Mycobacterial cell envelope

MCE 介导的脂质跨分枝杆菌细胞包膜转运的结构表征

• 批准号: 10620210
• 负责人: Casey Vieni
• 金额: $ 0.23万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2023-06-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10620210
• 关键词: Actinobacteria class; Adopted; Antibiotics; Architecture; Bacteria; Binding; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biological Models; Cause of Death; Cell membrane; Cell surface; Cells; Cholesterol; Communicable Diseases; Complement; Complex; Cryoelectron Microscopy; Cytoplasm; DNA Binding; DNA Sequence; DNA mapping; Drug Delivery Systems; Drug resistance; Environment; Escherichia coli; Gene Cluster; Genes; Genetic; Genetic Transcription; Genome; Genus Mycobacterium; Glycolipids; Hydrophobicity; Immune response; Knock-out; Lead; Life Cycle Stages; Lipids; Maintenance; Mammalian Cell; Mediating; Membrane; Molecular; Movement; Mutation; Mutation Analysis; Mycobacterium smegmatis; Mycobacterium tuberculosis; Mycobacterium tuberculosis complex; Mycolic Acid; Nutrient; Operon; Organism; Pathogenicity; Phenotype; Protein Family; Proteins; Regulation; Resources; Role; Specificity; Structure; System; Testing; Tuberculosis; Virulence; Virulence Factors; Work; antimicrobial; cell envelope; design; differential expression; improved; insight; lipid transport; mutant; mycobacterial; new therapeutic target; novel therapeutics; protein complex; rational design; resistant strain; response; structural biology; sulfolipids; tuberculosis treatment; uptake

PROJECT SUMMARY / ABSTRACT: The cell envelope of Mycobacterium tuberculosis (Mtb) serves an important role as protection against common antimicrobials and host cell defenses, but also a barrier through which Mtb must transport host derived nutrients. The MCE protein family (originally implicated in Mammalian Cell Entry) is nearly ubiquitous in double-membraned bacteria, and MCE proteins are now believed to act as transporters that facilitate lipid movement between the inner and outer membranes. While Mtb MCE proteins are critical for virulence, their mechanism in Mtb remains unclear. Due to Mtb’s distinct cell envelope Mtb MCE proteins likely adopt a unique architecture to mediate transport. In addition, throughout the course of Mtb infection MCE transporters are believed to be differentially expressed by MCE transcriptional regulators (MceRs). This suggests that environmental sensing in Mycobacteria in turn alters transcription of the mce operons. Using structural biology, biochemistry, and genetic complementation assays, I will provide the first glimpse into the architecture of Mtb Mce protein systems and elucidate how MceRs interact with their operator sequences at the molecular level to regulate mce operons. To do so I propose the following aims: (Aim 1) structure determination of the Mce4 lipid transport system from Mycobacterium smegmatis and (Aim 2) structural characterization and DNA-binding specificity of MceRs. My work will provide the first glimpse at the architecture of the M. smegmatis Mce4 transporter system, as well as the molecular mechanisms of MceR mediated mce operon regulation, which may shed light on the mechanisms of cell envelope maintenance in Mtb and provide a novel target for therapeutics.

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