A chimeric lectin formed from Bauhinia purpurea lectin and Lens culinaris lectin recognizes a unique carbohydrate structure.
Lectins are carbohydrate-binding proteins widely used in biochemical, immunochemical, and histochemical studies. Bauhinia purpurea lectin (BPA) is a leguminous lectin with an affinity for galactose and lactose. Nine amino acids, DTWPNTEWS, corresponding to the amino acid sequence from aspartic acid-135 to serine-143 in the primary structure of BPA were replaced with the corresponding amino acid residues from the mannose-binding Lens culinaris lectin (LCA), and the chimeric lectin obtained was expressed in Escherichia coli cells. The carbohydrate-binding specificity of the recombinant chimeric lectin was investigated in detail by comparing the elution profiles of various glycopeptides and oligosaccharides with defined carbohydate structures from immobilized lectin columns. Glycopeptides carrying three constitutive carbohydrate sequences of Galbeta1-3GalNAc-Ser/Thr and a complex-type biantennary glycopeptide, which show a high affinity for BPA or LCA, were shown to have no affinity for the chimeric lectin. In contrast, hybrid-type and high mannose-type glycopeptides with a Manalpha1-6(Manalpha1-3)Manalpha1-6Man sequence were found to have a moderate affinity for the chimeric lectin. This result demonstrates that a novel type of lectin with a unique carbohydrate-binding specificity can be constructed from BPA by substituting several amino acid residues in its metal-binding region with other amino acid residues. Additional lectin(s) with distinctly different carbohydrate-binding specificities will provide a powerful tool for many studies.
Yamamoto
K
K
Laboratory of Molecular Medicine, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. yamamoto@k.u-tokyo. ac.jp
Konami
Y
Y
Osawa
T
T
D002240:Carbohydrate Sequence; D007887:Fabaceae / Q000737:chemistry / Q000235:genetics*; D006020:Glycopeptides / Q000235:genetics / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D009844:Oligosaccharides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D037121:Plant Lectins; D010940:Plant Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism; D010946:Plants, Medicinal; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000378:metabolism*
C039340:Bauhinia purpurea lectin; D006020:Glycopeptides; D037102:Lectins; D009842:Oligopeptides; D009844:Oligosaccharides; D037121:Plant Lectins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C033619:lentil lectin