Structure determination of CAMP factor of Mobiluncus curtisii and insights into structural dynamics

Int J Biol Macromol. 2020 May 1:150:1027-1036. doi: 10.1016/j.ijbiomac.2019.10.107. Epub 2019 Nov 15.

Abstract

Bacterial vaginosis (BV) is a common type of vaginal inflammation caused by a proliferation of pathogenic bacteria, among which Mobiluncus curtisii. In our previous studies on M. curtisii genome, we identified the presence of a genomic fragment encoding a 25 kDa pore-forming toxin, the CAMP factor, which is known to be involved in the synergistic lysis of erythrocytes namely CAMP reaction. However, whether this hypothetical gene product has hemolytic activity is unknown. Moreover, its relative structure and function are not yet solved. Here we found that the M. curtisii CAMP factor is a monomer at pH 4.4 and oligomer at pH > 4.6. Hemolysis assays showed that M. curtisii CAMP factor could lyse sheep red blood cells efficiently in pH 5.4-7.4. Negative staining electron microscope analysis of the CAMP factor revealed ring-like structures at pH above 4.6. Additionally, the crystal structure of M. curtisii CAMP factor, determineded at 1.85 Å resolution, reveals a 5 + 3 helix motif. Further functional analysis suggested that the structural rearrangement of the N-terminal domain might be required for protein function. In conclusion, this structure-function relationship study of CAMP factor provides a new perspective of the M. curtisii role in BV development.

Keywords: CAMP factor; Conformational change; Hemolytic reaction; Mobiluncus curtisii; Pore-forming toxin; Structure determination.

MeSH terms

  • Actinomycetales Infections / genetics
  • Actinomycetales Infections / metabolism
  • Animals
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Erythrocytes / metabolism
  • Erythrocytes / microbiology
  • Female
  • Humans
  • Mobiluncus / chemistry*
  • Mobiluncus / genetics
  • Mobiluncus / metabolism
  • Molecular Dynamics Simulation*
  • Pore Forming Cytotoxic Proteins / chemistry*
  • Pore Forming Cytotoxic Proteins / genetics
  • Pore Forming Cytotoxic Proteins / metabolism
  • Protein Domains
  • Sheep
  • Structure-Activity Relationship
  • Vaginosis, Bacterial / genetics
  • Vaginosis, Bacterial / metabolism

Substances

  • Bacterial Proteins
  • Pore Forming Cytotoxic Proteins