DOI: 10.1073/pnas.1720996115
论文题名: Imaging mycobacterial growth and division with a fluorogenic probe
作者: Hodges H.L. ; Brown R.A. ; Crooks J.A. ; Weibel D.B. ; Kiessling L.L.
刊名: Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
出版年: 2018
卷: 115, 期: 20 起始页码: 5271
结束页码: 5276
语种: 英语
英文关键词: Ag85
; Cell wall
; Lipid
; Mycolic acid
; Tuberculosis
Scopus关键词: antibiotic agent
; peptidoglycan
; quencher trehalose fluorophore
; transferase
; unclassified drug
; bacterial protein
; cord factor
; fluorescent dye
; peptidoglycan
; trehalose monomycolate
; Article
; bacterial growth
; bacterial membrane
; bacterial phenomena and functions
; bacterium division
; bilayer membrane
; cell division
; cellular distribution
; complex formation
; controlled study
; enzyme activity
; enzyme substrate
; extracellular space
; fluorescence analysis
; host resistance
; hydrolysis
; image analysis
; intracellular membrane
; live cell imaging
; molecular probe
; Mycobacterium
; nonhuman
; priority journal
; taxonomic rank
; transesterification
; cell wall
; chemistry
; Corynebacterium glutamicum
; fluorescence
; growth, development and aging
; human
; image processing
; metabolism
; microbiology
; Mycobacterium tuberculosis
; procedures
; tuberculosis
; Bacterial Proteins
; Cell Division
; Cell Wall
; Cord Factors
; Corynebacterium glutamicum
; Fluorescence
; Fluorescent Dyes
; Humans
; Image Processing, Computer-Assisted
; Mycobacterium tuberculosis
; Peptidoglycan
; Tuberculosis
英文摘要: Control and manipulation of bacterial populations requires an understanding of the factors that govern growth, division, and antibiotic action. Fluorescent and chemically reactive small molecule probes of cell envelope components can visualize these processes and advance our knowledge of cell envelope biosynthesis (e.g., peptidoglycan production). Still, fundamental gaps remain in our understanding of the spatial and temporal dynamics of cell envelope assembly. Previously described reporters require steps that limit their use to static imaging. Probes that can be used for real-time imaging would advance our understanding of cell envelope construction. To this end, we synthesized a fluorogenic probe that enables continuous live cell imaging in mycobacteria and related genera. This probe reports on the mycolyltransferases that assemble the mycolic acid membrane. This peptidoglycan-anchored bilayer-like assembly functions to protect these cells from antibiotics and host defenses. Our probe, quencher-trehalose-fluorophore (QTF), is an analog of the natural mycolyltransferase substrate. Mycolyltransferases process QTF by diverting their normal transesterification activity to hydrolysis, a process that unleashes fluorescence. QTF enables high contrast continuous imaging and the visualization of mycolyltransferase activity in cells. QTF revealed that mycolyltransferase activity is augmented before cell division and localized to the septa and cell poles, especially at the old pole. This observed localization suggests that mycolyltransferases are components of extracellular cell envelope assemblies, in analogy to the intracellular divisomes and polar elongation complexes. We anticipate QTF can be exploited to detect and monitor mycobacteria in physiologically relevant environments. © 2018 National Academy of Sciences. All rights reserved. Citation statistics:
资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/163714
Appears in Collections: 气候变化与战略
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作者单位: Hodges, H.L., Department of Chemistry, University of Wisconsin–Madison, Madison, WI 53706, United States; Brown, R.A., Department of Chemistry, University of Wisconsin–Madison, Madison, WI 53706, United States; Crooks, J.A., Department of Biochemistry, University of Wisconsin–Madison, Madison, WI 53706, United States; Weibel, D.B., Department of Biochemistry, University of Wisconsin–Madison, Madison, WI 53706, United States; Kiessling, L.L., Department of Chemistry, University of Wisconsin–Madison, Madison, WI 53706, United States, Department of Biochemistry, University of Wisconsin–Madison, Madison, WI 53706, United States, Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States
Recommended Citation:
Hodges H.L.,Brown R.A.,Crooks J.A.,et al. Imaging mycobacterial growth and division with a fluorogenic probe[J]. Proceedings of the National Academy of Sciences of the United States of America,2018-01-01,115(20)