Influence of Impaired Lipoprotein Biogenesis on Surface and Exoproteome of Streptococcus Pneumoniae

J Proteome Res. 2014 Feb 7;13(2):650-67. doi: 10.1021/pr400768v. Epub 2014 Jan 15.

Abstract

Surface proteins are important for the fitness and virulence of the Gram-positive pathogen Streptococcus pneumoniae. They are crucial for interaction of the pathogen with its human host during infection. Therefore, the analysis of the pneumococcal surface proteome is an important task that requires powerful tools. In this study, two different methods, an optimized biotinylation approach and shaving with trypsin beads, were applied to study the pneumococcal surface proteome and to identify surface-exposed protein domains, respectively. The identification of nearly 95% of the predicted lipoproteins and 75% of the predicted sortase substrates reflects the high coverage of the two classical surface protein classes accomplished in this study. Furthermore, the biotinylation approach was applied to study the impact of an impaired lipoprotein maturation pathway on the cell envelope proteome and exoproteome. Loss of the lipoprotein diacylglyceryl transferase Lgt leads to striking changes in the lipoprotein distribution. Many lipoproteins disappear from the surface proteome and accumulate in the exoproteome. Further insights into lipoprotein processing in pneumococci are provided by immunoblot analyses of bacterial lysates and corresponding supernatant fractions. Taken together, the first comprehensive overview of the pneumococcal surface and exoproteome is presented, and a model for lipoprotein processing in S. pneumoniae is proposed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / metabolism
  • Base Sequence
  • Biotin / metabolism
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Lipoproteins / biosynthesis*
  • Lipoproteins / metabolism
  • Polymerase Chain Reaction
  • Proteome*
  • Streptococcus pneumoniae / metabolism*
  • Subcellular Fractions / metabolism
  • Trypsin / metabolism

Substances

  • Bacterial Proteins
  • DNA Primers
  • Lipoproteins
  • Proteome
  • Biotin
  • Trypsin