Mannose binding lectin and lung collectins interact with Toll-like receptor 4 and MD-2 by different mechanisms

• Published in: Biochimica et biophysica acta Vol. 1790; no. 12; pp. 1705 - 1710
• Main Authors: Shimizu, Takeyuki, Nishitani, Chiaki, Mitsuzawa, Hiroaki, Ariki, Shigeru, Takahashi, Motoko, Ohtani, Katsuki, Wakamiya, Nobutaka, Kuroki, Yoshio
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2009
• Subjects: Animals; Cells, Cultured; Collectin; Collectins - metabolism; Humans; Innate immunity; Lung - metabolism; Lymphocyte Antigen 96 - metabolism; Mannose binding lectin; Mannose-Binding Lectin - metabolism; MD-2; Protein Binding; Rats; Recombinant Proteins - metabolism; Signal Transduction - physiology; Spodoptera; Surfactant protein; Toll-like receptor 4; Toll-Like Receptor 4 - metabolism; Surfactant protein; MD-2; Innate immunity; Collectin; Mannose binding lectin; Toll-like receptor 4
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2009.10.006

We have previously shown that lung collectins, surfactant protein A (SP-A) and surfactant protein D, interact with Toll-like receptor (TLR) 2, TLR4, or MD-2. Bindings of lung collectins to TLR2 and TLR4/MD-2 result in the alterations of signaling through these receptors, suggesting the immunomodulatory functions of lung collectins. Mannose binding lectin (MBL) is another collectin molecule which has structural homology to SP-A. The interaction between MBL and TLRs has not yet been determined. We prepared recombinant MBL, and analyzed its bindings to recombinant soluble forms of TLR4 (sTLR4) and MD-2. MBL bound to sTLR4 and MD-2. The interactions were Ca 2+-dependent and inhibited by mannose or monoclonal antibody against the carbohydrate-recognition domain of MBL. Treatment of sTLR4 or MD-2 by peptide N-glycosidase F significantly decreased the binding of MBL. SP-A bound to deglycosylated sTLR4, and this property did not change in chimeric molecules of SP-A/MBL in which Glu 195–Phe 228 or Thr 174–Gly 194 of SP-A were replaced with the corresponding MBL sequences. These results suggested that MBL binds to TLR4 and MD-2 through the carbohydrate-recognition domain, and that oligosaccharide moieties of TLR4 and MD-2 are important for recognition by MBL. Since our previous studies indicated that lung collectins bind to the peptide portions of TLRs, MBL and lung collectins interact with TLRs by different mechanisms. These direct interactions between MBL and TLR4 or MD-2 suggest that MBL may modulate cellular responses by altering signals through TLRs.