title: Nanoscopic analysis on pH induced morphological changes of flagella in Escherichia coli.
abstract: OBJECTIVE
  Flagella contribute to the virulence of pathogenic bacteria through chemotaxis, motility, and adhesion. Understanding the various functions of flagella may provide insight into mechanisms of bacterial infection and transmission. The objectives of our study were to apply biophysical and biochemical methods to investigate the mechanisms of pH-dependent changes in flagella functions.
  
  
  METHODS
  Atomic force microscopy (AFM) was used to analyze the flagellum morphology of Escherichia coli cultured in various pH conditions. The swarming plate method was used to identify pH-dependent changes in bacterial motility. Western blot analysis and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) were also carried out to study pH-dependent expression and structural changes of flagellin C.
  
  
  RESULTS
  E coli cultured at pH 7 produced the flagella with the greatest average length and diameter. When the bacteria were grown at pH 6 or pH 8, shorter and thinner forms of flagella were produced. The morphology of the flagella was correlated to the bacterial motility. While western blot analysis showed only a slight change in the expression of the flagellin C protein in response to changes in the pH of the culture medium, ATR-FTIR showed significant pH-dependent changes in the secondary structure of the flagellin C assembled in sheared flagella.
  
  
  CONCLUSIONS
  Our results show that both acidification and alkalization of the culture medium restricted bacterial motility, and indicate that the reduced motility may be caused by incorrect assembly of the flagellum proteins.
authors.affiliation: Graduate Institute of Medical Biotechnology, Tzu Chi University, Hualien 970, Taiwan.
authors.forename: Kai-Chih
authors.initials: KC
authors.lastname: Chang
authors.affiliation: Graduate Institute of Medical Biotechnology, Tzu Chi University, Hualien 970, Taiwan.
authors.forename: Shun-Jia
authors.initials: SJ
authors.lastname: Cheng
authors.affiliation: Department of Medicine, Mackay Medical College, New Taipei 252, Taiwan.
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Chen
authors.affiliation: Department of Biochemistry, School of Medicine, Tzu Chi University, Hualien 970, Taiwan.
authors.forename: Hsin-Ru
authors.initials: HR
authors.lastname: Huang
authors.affiliation: Graduate Institute of Medical Biotechnology, Tzu Chi University, Hualien 970, Taiwan; Department of Biochemistry, School of Medicine, Tzu Chi University, Hualien 970, Taiwan. Electronic address: jwliou@mail.tcu.edu.tw.
authors.forename: Je-Wen
authors.initials: JW
authors.lastname: Liou
mesh_terms: D001669:Biochemical Phenomena / Q000187:drug effects; D055592:Biophysical Phenomena / Q000187:drug effects; D015153:Blotting, Western; D004926:Escherichia coli / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D005407:Flagella / Q000502:physiology* / Q000648:ultrastructure; D005408:Flagellin / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008124:Locomotion; D018625:Microscopy, Atomic Force; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D005408:Flagellin