Solution NMR studies provide structural basis for endotoxin pattern recognition by the innate immune receptor CD14.
CD14 functions as a key pattern recognition receptor for a diverse array of Gram-negative and Gram-positive cell-wall components in the host innate immune response by binding to pathogen-associated molecular patterns (PAMPs) at partially overlapping binding site(s). To determine the potential contribution of CD14 residues in this pattern recognition, we have examined using solution NMR spectroscopy, the binding of three different endotoxin ligands, lipopolysaccharide, lipoteichoic acid, and a PGN-derived compound, muramyl dipeptide to a 15N isotopically labeled 152-residue N-terminal fragment of sCD14 expressed in Pichia pastoris. Mapping of NMR spectral changes upon addition of ligands revealed that the pattern of residues affected by binding of each ligand is partially similar and partially different. This first direct structural observation of the ability of specific residue combinations of CD14 to differentially affect endotoxin binding may help explain the broad specificity of CD14 in ligand recognition and provide a structural basis for pattern recognition. Another interesting finding from the observed spectral changes is that the mode of binding may be dynamically modulated and could provide a mechanism for binding endotoxins with structural diversity through a common binding site.
Albright
Seth
S
Biochemistry, Cellular and Molecular Biology Department, University of Tennessee, M407 Walters Life Sciences, 1410 Cumberland Avenue, Knoxville, TN 37996-0840, USA.
Chen
Bin
B
Holbrook
Kristen
K
Jain
Nitin U
NU
D000937:Antigen-Antibody Reactions / Q000276:immunology; D001665:Binding Sites; D004731:Endotoxins / Q000737:chemistry* / Q000276:immunology*; D007113:Immunity, Innate / Q000276:immunology*; D018950:Lipopolysaccharide Receptors / Q000737:chemistry* / Q000276:immunology*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D046709:Pattern Recognition, Physiological / Q000502:physiology*; D010843:Pichia / Q000276:immunology*; D011485:Protein Binding; D013329:Structure-Activity Relationship
D004731:Endotoxins; D018950:Lipopolysaccharide Receptors