RRC ID |
1288
|
Author |
Adachi Y, Ishii T, Ikeda Y, Hoshino A, Tamura H, Aketagawa J, Tanaka S, Ohno N.
|
Title |
Characterization of beta-glucan recognition site on C-type lectin, dectin 1.
|
Journal |
Infect Immun
|
Abstract |
Dectin 1 is a mammalian cell surface receptor for (1-->3)-beta-d-glucans. Since (1-->3)-beta-d-glucans are commonly present on fungal cell walls, it has been suggested that dectin 1 is important for recognizing fungal invasion. In this study we tried to deduce the amino acid residues in dectin 1 responsible for beta-glucan recognition. HEK293 cells transfected with mouse dectin 1 cDNA could bind to a gel-forming (1-->3)-beta-d-glucan, schizophyllan (SPG). The binding of SPG to a dectin 1 transfectant was inhibited by pretreatment with other beta-glucans having a (1-->3)-beta-d-glucosyl linkage but not by pretreatment with alpha-glucans. Dectin 1 has a carbohydrate recognition domain (CRD) consisting of six cysteine residues that are highly conserved in C-type lectins. We prepared 32 point mutants with mutations in the CRD and analyzed their binding to SPG. Mutations at Trp(221) and His(223) resulted in decreased binding to beta-glucan. Monoclonal antibody 4B2, a dectin- 1 monoclonal antibody which had a blocking effect on the beta-glucan interaction, completely failed to bind the dectin-1 mutant W221A. A mutant with mutations in Trp(221) and His(223) did not have a collaborative effect on Toll-like receptor 2-mediated cellular activation in response to zymosan. These amino acid residues are distinct from residues in other sugar-recognizing peptide sequences of typical C-type lectins. These results suggest that the amino acid sequence W221-I222-H223 is critical for formation of a beta-glucan binding site in the CRD of dectin 1.
|
Volume |
72(7)
|
Pages |
4159-71
|
Published |
2004-7-1
|
DOI |
10.1128/IAI.72.7.4159-4171.2004
|
PII |
72/7/4159
|
PMID |
15213161
|
PMC |
PMC427417
|
MeSH |
Amino Acid Sequence
Animals
Antibodies, Monoclonal / immunology
Binding Sites
Biotin / metabolism
Glucans / chemistry
Glucans / metabolism*
Lectins, C-Type / chemistry
Lectins, C-Type / metabolism*
Membrane Proteins / chemistry
Membrane Proteins / genetics
Membrane Proteins / immunology
Membrane Proteins / metabolism*
Mice
Molecular Sequence Data
Mutation
NF-kappa B / metabolism
Nerve Tissue Proteins / chemistry
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / immunology
Nerve Tissue Proteins / metabolism*
Protein Structure, Tertiary
Schizophyllum / genetics
Schizophyllum / immunology
Schizophyllum / metabolism
Sequence Analysis, Protein
|
IF |
3.201
|
Times Cited |
154
|
WOS Category
|
INFECTIOUS DISEASES
IMMUNOLOGY
|
Resource |
Human and Animal Cells |
RAW 264(RCB0535) |