The bactericidal activity of the C-type lectin RegIIIβ against Gram-negative bacteria involves binding to lipid A.
RegIIIβ is a member of the C-type lectin family called RegIII. It is known to bind peptidoglycan, and its bactericidal activity shapes the interactions with commensal and pathogenic gut bacteria. However, little is known about its carbohydrate recognition specificity and the bactericidal mechanism, particularly against Gram-negative bacteria. Here, we show that RegIIIβ can bind directly to LPS by recognizing the carbohydrate moiety of lipid A via a novel motif that is indispensable for its bactericidal activity. This bactericidal activity of RegIIIβ could be inhibited by preincubation with LPS, lipid A, or gentiobiose. The latter is a disaccharide composed of two units of β-(1→6)-linked d-glucose and resembles the carbohydrate moiety of lipid A. Therefore, this structural element may form a key target site recognized by RegIIIβ. Using point-mutated RegIIIβ proteins, we found that amino acid residues in two structural motifs termed "loop 1" and "loop 2," are important for peptidoglycan and lipid A binding (Arg-135, Asp-142) and for the bactericidal activity (Glu-134, Asn-136, Asp-142). Thus, the ERN motif and residue Asp-142 in the loop 2 are of critical importance for RegIIIβ function. This provides novel insights into the carbohydrate recognition specificity of RegIIIβ and explains its bactericidal activity against Gram-negative bacteria.
Institute of Microbiology, Department of Biology, ETH Zürich, 8093 Zürich, Switzerland.
Tsuyoshi
T
Miki
Otto
O
Holst
Wolf-Dietrich
WD
Hardt
D020816:Amino Acid Motifs; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism; D004187:Disaccharides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000737:chemistry*; D037181:Lectins, C-Type / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008050:Lipid A / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D000073718:Pancreatitis-Associated Proteins; D017354:Point Mutation; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D000900:Anti-Bacterial Agents; D004187:Disaccharides; D037181:Lectins, C-Type; D008050:Lipid A; D000073718:Pancreatitis-Associated Proteins; D011506:Proteins; C490253:Reg3b protein, mouse; C100052:gentiobiose