TY - JOUR
T1 - Identification of staphylococcal lipoteichoic acid-binding proteins in human serum by high-resolution LTQ-Orbitrap mass spectrometry
AU - Jang, Kyoung Soon
AU - Baik, Jung Eun
AU - Kang, Seok Seong
AU - Jeon, Jun Ho
AU - Choi, Seulggie
AU - Yang, Yung Hun
AU - Kim, Byung Gee
AU - Yun, Cheol Heui
AU - Han, Seung Hyun
PY - 2012/3
Y1 - 2012/3
N2 - 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.
AB - 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.
KW - Lipoteichoic acid
KW - LTA-binding protein
KW - LTQ-Orbitrap mass spectrometry
KW - Staphylococcus aureus
UR - http://www.scopus.com/inward/record.url?scp=84857368791&partnerID=8YFLogxK
U2 - 10.1016/j.molimm.2011.11.012
DO - 10.1016/j.molimm.2011.11.012
M3 - Article
C2 - 22189407
AN - SCOPUS:84857368791
SN - 0161-5890
VL - 50
SP - 177
EP - 183
JO - Molecular Immunology
JF - Molecular Immunology
IS - 3
ER -