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Related Experiment Videos

Characterization of a dynamic S layer on Bacillus thuringiensis.

M D Luckevich1, T J Beveridge

  • 1Department of Microbiology, College of Biological Science, University of Guelph, Ontario, Canada.

Journal of Bacteriology
|December 1, 1989
PubMed
Summary

This study characterizes the dynamic S layer of Bacillus thuringiensis, revealing a protein structure that is shed during cell growth and eventually lost, making it atypical and challenging to analyze.

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Area of Science:

  • Microbiology
  • Structural Biology
  • Biochemistry

Background:

  • Bacillus thuringiensis strains feature surface S layers, which are proteinaceous crystalline arrays.
  • S layers contribute to cell envelope structure and interactions.

Purpose of the Study:

  • To characterize the S layer of three Bacillus thuringiensis strains.
  • To investigate the S layer's structure, composition, and dynamic behavior during cell culture.

Main Methods:

  • Electron microscopy (platinum shadowing) for structural analysis.
  • Chemical extraction (guanidine hydrochloride) for protein isolation.
  • Biochemical analysis (pI, residue composition) and N-terminal sequencing.
  • Antibody labeling and immunoblotting to assess S layer abundance and integrity.

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  • Enzymatic assays (autolytic activity, protease digestion) to study S layer turnover.

    • Main Results:

    • The S layer exhibits p2 symmetry with specific lattice parameters (a=8.5 nm, b=7.2 nm, γ=73°).
    • The S layer protein is a hydrophilic 91.4-kDa molecule with a pI of 5, lacking certain amino acids and carbohydrates.
    • S layer shedding and degradation were observed, correlating with cell growth phase and autolytic/proteolytic activities.
    • S layer presence decreased with serial subculturing, indicating instability.

    Conclusions:

    • The Bacillus thuringiensis S layer is a dynamic structure, atypical compared to stable S layers in other bacteria.
    • Cellular processes like wall turnover and enzymatic activities contribute to S layer instability.
    • The dynamic nature complicates S layer characterization but offers insights into bacterial surface remodeling.