Identification of staphylococcal lipoteichoic acid-binding proteins in human serum by high-resolution LTQ-Orbitrap mass spectrometry

Kyoung Soon Jang, Jung Eun Baik, Seok Seong Kang, Jun Ho Jeon, Seulggie Choi, Yung Hun Yang, Byung Gee Kim, Cheol Heui Yun, Seung Hyun Han

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Lipoteichoic acid (LTA), a major virulence factor of Gram-positive bacteria, is associated with bacterial adherence to host cells, biofilm formation, and inflammation. LTA-binding proteins (LTA-BPs) play an important role in the host immune response by initially recognizing and responding to LTA during infections. In this study, we screened for LTA-BPs in human serum using LTA-immobilized beads and high-throughput mass spectrometry. Highly pure and structurally intact LTA was prepared from Staphylococcus aureus and immobilized onto N-hydroxysuccinimide-activated Sepharose ® 4 Fast Flow beads. The immobilization process does not seem to affect the biological activity of LTA since LTA-immobilized beads could stimulate macrophages and activate Toll-like receptor 2. Then, the LTA-immobilized beads were incubated with the human serum to capture LTA-BPs and their molecular identities were determined using high-resolution LTQ-Orbitrap hybrid Fourier transform mass spectrometry. LTA-BPs captured at high frequencies were neutrophil-activating peptide 2, prohibitin-2, alpha-1-anti-trypsin, histidine-rich glycoprotein, apolipoproteins, complements, and coagulation factor, most of which are known to be related with the host immune responses against infections. As high-throughput, efficient, accurate and sensitive, this screening method could be widely applicable to the identification of novel binding proteins to microbial virulence factors with glycolipid structures.

Original languageEnglish
Pages (from-to)177-183
Number of pages7
JournalMolecular Immunology
Volume50
Issue number3
DOIs
StatePublished - Mar 2012

Keywords

  • Lipoteichoic acid
  • LTA-binding protein
  • LTQ-Orbitrap mass spectrometry
  • Staphylococcus aureus

Fingerprint

Dive into the research topics of 'Identification of staphylococcal lipoteichoic acid-binding proteins in human serum by high-resolution LTQ-Orbitrap mass spectrometry'. Together they form a unique fingerprint.

Cite this