1. Characterization of two novel LPS-binding sites in leukocyte integrin betaA domain.
- Author
-
Wong KF, Luk JM, Cheng RH, Klickstein LB, and Fan ST
- Subjects
- Amino Acid Sequence, Animals, Binding Sites, CD18 Antigens chemistry, CD18 Antigens genetics, Humans, Jurkat Cells, Models, Molecular, Molecular Sequence Data, NF-kappa B metabolism, Peptides genetics, Peptides metabolism, Point Mutation, Protein Structure, Secondary, Sequence Alignment, Signal Transduction physiology, Tumor Necrosis Factor-alpha genetics, Tumor Necrosis Factor-alpha metabolism, CD18 Antigens metabolism, Leukocytes immunology, Lipopolysaccharides metabolism
- Abstract
Lipopolysaccharide (LPS), a bacterial endotoxin, triggers deleterious systemic inflammatory responses when released into blood circulation, causing organ dysfunction and death. In response to LPS stimulation, CD14 and toll-like receptor (TLR)-4 elicit inflammatory signaling cascades. Although leukocyte integrins (CD11b/CD18 and CD11c/CD18) were reported to bind LPS and induce NF-kappaB translocation, the evidence on such epitope location remains elusive. The present study aims to delineate the LPS-binding sites on the integrin CD18 antigen and to design peptide(s) as potential prophylactic and/or therapeutic agents to modulate LPS effects in activated Jurkat cells. Epitope mapping analysis using a series of CD18 truncated variants revealed two putative LPS-binding sites within the betaA region (216-248 and 266-318 a.a.), which were further confirmed by point mutation studies. Inhibition assay demonstrated that the CD18-betaA(266-318) peptide could block LPS binding in a dose-dependent manner. Our data also indicated that treatment with the CD18-peptide modulated TNF-alpha mRNA transcription via the NF-kappaB signaling pathway in LPS-activated Jurkat cells. In conclusion, we have identified two novel LPS-binding sites located at the CD18 betaA domain of leukocyte integrin, and the integrin peptide betaA(266-318) is shown to inhibit LPS binding and subsequent inflammatory events, having therapeutic implications to cure gram-negative endotoxemia.
- Published
- 2007
- Full Text
- View/download PDF