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

Research output: Contribution to journalArticle

44 Citations (Scopus)

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

Fingerprint

Toll-Like Receptor 2
Lipoproteins
Staphylococcus aureus
Lipopeptides
High pressure liquid chromatography
Deoxycholic Acid
Trypsin
Glass
Polyacrylamide Gel Electrophoresis
Digestion
High Pressure Liquid Chromatography
Cells
Ligands
Peptides
Molecules

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

Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus. / Tawaratsumida, Kazuki; Furuyashiki, Maiko; Katsumoto, Mami; Fujimoto, Yukari; Fukase, Koichi; Suda, Yasuo; Hashimoto, Masahito.

In: Journal of Biological Chemistry, Vol. 284, No. 14, 03.04.2009, p. 9147-9152.

Research output: Contribution to journalArticle

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, vol. 284, no. 14, pp. 9147-9152. https://doi.org/10.1074/jbc.M900429200
Tawaratsumida, Kazuki ; Furuyashiki, Maiko ; Katsumoto, Mami ; Fujimoto, Yukari ; Fukase, Koichi ; Suda, Yasuo ; Hashimoto, Masahito. / Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 14. pp. 9147-9152.
@article{76c33dd707ec4ea194f6266191fa930e,
title = "Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus",
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.",
author = "Kazuki Tawaratsumida and Maiko Furuyashiki and Mami Katsumoto and Yukari Fujimoto and Koichi Fukase and Yasuo Suda and Masahito Hashimoto",
year = "2009",
month = "4",
day = "3",
doi = "10.1074/jbc.M900429200",
language = "English",
volume = "284",
pages = "9147--9152",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus

AU - Tawaratsumida, Kazuki

AU - Furuyashiki, Maiko

AU - Katsumoto, Mami

AU - Fujimoto, Yukari

AU - Fukase, Koichi

AU - Suda, Yasuo

AU - Hashimoto, Masahito

PY - 2009/4/3

Y1 - 2009/4/3

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=66149125243&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=66149125243&partnerID=8YFLogxK

U2 - 10.1074/jbc.M900429200

DO - 10.1074/jbc.M900429200

M3 - Article

C2 - 19218237

AN - SCOPUS:66149125243

VL - 284

SP - 9147

EP - 9152

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -