Characteristic recognition of N-acetylgalactosamine by an invertebrate C-type Lectin, CEL-I, revealed by X-ray crystallographic analysis.
CEL-I is a C-type lectin, purified from the sea cucumber Cucumaria echinata, that shows a high specificity for N-acetylgalactosamine (GalNAc). We determined the crystal structures of CEL-I and its complex with GalNAc at 2.0 and 1.7 A resolution, respectively. CEL-I forms a disulfide-linked homodimer and contains two intramolecular disulfide bonds, although it lacks one intramolecular disulfide bond that is widely conserved among various C-type carbohydrate recognition domains (CRDs). Although the sequence similarity of CEL-I with other C-type CRDs is low, the overall folding of CEL-I was quite similar to those of other C-type CRDs. The structure of the complex with GalNAc revealed that the basic recognition mode of GalNAc was very similar to that for the GalNAc-binding mutant of the mannose-binding protein. However, the acetamido group of GalNAc appeared to be recognized more strongly by the combination of hydrogen bonds to Arg115 and van der Waals interaction with Gln70. Mutational analyses, in which Gln70 and/or Arg115 were replaced by alanine, confirmed that these residues contributed to GalNAc recognition in a cooperative manner.
Research Center for Structural and Functional Proteomics, Institute for Protein Research, Osaka University, 3-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
Hajime
H
Sugawara
Masami
M
Kusunoki
Genji
G
Kurisu
Tokiko
T
Fujimoto
Haruhiko
H
Aoyagi
Tomomitsu
T
Hatakeyama
D000116:Acetylgalactosamine / Q000737:chemistry*; D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry; D002118:Calcium / Q000737:chemistry; D002241:Carbohydrates / Q000737:chemistry; D018360:Crystallography, X-Ray; D048016:Cucumaria; D004252:DNA Mutational Analysis; D019281:Dimerization; D004220:Disulfides; D005973:Glutamine / Q000737:chemistry; D006860:Hydrogen Bonding; D037181:Lectins, C-Type / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid
D002241:Carbohydrates; D004220:Disulfides; D037181:Lectins, C-Type; D011994:Recombinant Proteins; D005973:Glutamine; D001120:Arginine; D000116:Acetylgalactosamine; D000409:Alanine; D002118:Calcium