Autocatalytic association of proteins by covalent bond formation: a Bio Molecular Welding toolbox derived from a bacterial adhesin

Sci Rep. 2017 Mar 2;7:43564. doi: 10.1038/srep43564.

Abstract

Unusual intramolecular cross-links present in adhesins from Gram-positive bacteria have been used to develop a generic process amenable to biotechnology applications. Based on the crystal structure of RrgA, the Streptococcus pneumoniae pilus adhesin, we provide evidence that two engineered protein fragments retain their ability to associate covalently with high specificity, in vivo and in vitro, once isolated from the parent protein. We determined the optimal conditions for the assembly of the complex and we solved its crystal structure at 2 Å. Furthermore, we demonstrate biotechnological applications related to antibody production, nanoassembly and cell-surface labeling based on this process we named Bio Molecular Welding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / chemistry*
  • Adhesins, Bacterial / genetics
  • Adhesins, Bacterial / metabolism
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Catalysis
  • Fimbriae Proteins / chemistry
  • Fimbriae Proteins / genetics
  • Fimbriae Proteins / metabolism
  • Models, Molecular
  • Molecular Weight
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / metabolism
  • Protein Binding
  • Protein Conformation
  • Recombinant Fusion Proteins
  • Spectrometry, Mass, Electrospray Ionization
  • Virulence Factors / chemistry
  • Virulence Factors / genetics
  • Virulence Factors / metabolism

Substances

  • Adhesins, Bacterial
  • Bacterial Proteins
  • Multiprotein Complexes
  • Recombinant Fusion Proteins
  • RrgA protein, Streptococcus pneumoniae
  • Virulence Factors
  • Fimbriae Proteins