Modulation of the Bacillus anthracis secretome by the immune inhibitor A1 protease

J Bacteriol. 2014 Jan;196(2):424-35. doi: 10.1128/JB.00690-13. Epub 2013 Nov 8.

Abstract

The Bacillus anthracis secretome includes protective antigen, lethal factor, and edema factor, which are the components of anthrax toxin, and other proteins with known or potential roles in anthrax disease. Immune inhibitor A1 (InhA1) is a secreted metalloprotease that is unique to pathogenic members of the Bacillus genus and has been associated with cleavage of host proteins during infection. Here, we report the effect of InhA1 on the B. anthracis secretome. Differential in-gel electrophoresis of proteins present in culture supernatants from a parent strain and an isogenic inhA1-null mutant revealed multiple differences. Of the 1,340 protein spots observed, approximately one-third were less abundant and one-third were more abundant in the inhA1 secretome than in the parent strain secretome. Proteases were strongly represented among those proteins exhibiting a 9-fold or greater change. InhA1 purified from a B. anthracis culture supernatant directly cleaved each of the anthrax toxin proteins as well as an additional secreted protease, Npr599. The conserved zinc binding motif HEXXH of InhA1 (HEYGH) was critical for its proteolytic activity. Our data reveal that InhA1 directly and indirectly modulates the form and/or abundance of over half of all the secreted proteins of B. anthracis. The proteolytic activity of InhA1 on established secreted virulence factors, additional proteases, and other secreted proteins suggests that this major protease plays an important role in virulence not only by cleaving mammalian substrates but also by modulating the B. anthracis secretome itself.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Motifs
  • Antigens, Bacterial / metabolism*
  • Bacillus anthracis / genetics
  • Bacillus anthracis / metabolism*
  • Bacterial Proteins / metabolism*
  • Bacterial Toxins / metabolism*
  • Binding Sites
  • Electrophoresis
  • Gene Deletion
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism*
  • Protein Binding
  • Proteome / analysis
  • Zinc / metabolism

Substances

  • Antigens, Bacterial
  • Bacterial Proteins
  • Bacterial Toxins
  • Proteome
  • anthrax toxin
  • Metalloendopeptidases
  • microbial metalloproteinases
  • Zinc