Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus

Kazuki Tawaratsumida, Maiko Furuyashiki, Mami Katsumoto, Yukari Fujimoto, Koichi Fukase, Yasuo Suda, Masahito Hashimoto

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Abstract

Staphylococcus aureus is known to activate mammalian immune cells through Toll-like receptor 2 (TLR2). We recently demonstrated that a lipoprotein fraction obtained from S. aureus by Triton X-114 phase partitioning is a potent activator of TLR2. In this study, we separated TLR2-activating lipoproteins expressed in S. aureus and characterized an N-terminal structure. The lipoprotein fraction of S. aureus was prepared by glass bead disruption followed by Triton X-114 phase partitioning. The TLR2-activating molecules were mainly detected in the mass range of 30-35 kDa. Seven lipoproteins were identified by the mass spectra of their tryptic digests. Among them, three lipoproteins were separated by preparative SDS-PAGE and proved to activate TLR2. After digestion with trypsin in the presence of sodium deoxycholate, the N terminus of the lipopeptide was isolated from lipoprotein SAOUHSC_02699 by normal phase high pressure liquid chromatography and characterized as an S-(diacyloxypropyl)cystein-containing peptide using tandem mass spectra. The synthetic lipopeptide counter-part also stimulated the cells via TLR2. These results showed that the diacylated lipoprotein from S. aureus acts as a TLR2 ligand in mammalian cells.

Original languageEnglish
Pages (from-to)9147-9152
Number of pages6
JournalJournal of Biological Chemistry
Volume284
Issue number14
DOIs
Publication statusPublished - 2009 Apr 3
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Tawaratsumida, K., Furuyashiki, M., Katsumoto, M., Fujimoto, Y., Fukase, K., Suda, Y., & Hashimoto, M. (2009). Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus. Journal of Biological Chemistry, 284(14), 9147-9152. https://doi.org/10.1074/jbc.M900429200