Konami Y, Yamamoto K, Osawa T, Irimura T
Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
FEBS Lett. 1992 Jun 15;304(2-3):129-35. doi: 10.1016/0014-5793(92)80603-e.
A carbohydrate-binding peptide of the di-N-acetylchitobiose-binding Cytisus sessilifolius anti-H(O) lectin I (CSA-I) was isolated from the endoproteinase Asp-N digest of CSA-I by affinity chromatography on a column of N-acetyl-D-glucosamine oligomer-Sepharose (GlcNAc oligomer-Sepharose). The amino acid sequence of the carbohydrate-binding peptide of CSA-I was determined to be DTYFGKTYNPW using a gas-phase protein sequencer. This sequence corresponds to the sequence from Asp-129 to Trp-139 based on the primary structure of CSA-I, and shows a high degree of homology to those of the putative carbohydrate-binding peptide of the Laburnum alpinum lectin I (LAA-I) (DTYFGKAYNPW) and of the Ulex europaeus lectin II (UEA-II) (DSYFGKTYNPW). The binding of these three anti-H(O) lectins is known to be inhibited by di-N-acetylchitobiose but not by L-fucose. These results strongly suggest that there is a good correlation between the carbohydrate-binding specificity and the amino acid sequence of the carbohydrate-binding regions of di-N-acetylchitobiose-binding lectins.
通过在N-乙酰-D-葡糖胺寡聚物-琼脂糖（GlcNAc寡聚物-琼脂糖）柱上进行亲和层析，从无柄金雀花抗H（O）凝集素I（CSA-I）的内肽酶Asp-N消化物中分离出二-N-乙酰壳二糖结合的碳水化合物结合肽。使用气相蛋白质测序仪确定CSA-I的碳水化合物结合肽的氨基酸序列为DTYFGKTYNPW。基于CSA-I的一级结构，该序列对应于从Asp-129到Trp-139的序列，并且与高山豆凝集素I（LAA-I）（DTYFGKAYNPW）和荆豆凝集素II（UEA-II）（DSYFGKTYNPW）的推定碳水化合物结合肽的序列具有高度同源性。已知这三种抗H（O）凝集素的结合可被二-N-乙酰壳二糖抑制，但不能被L-岩藻糖抑制。这些结果强烈表明，二-N-乙酰壳二糖结合凝集素的碳水化合物结合特异性与其碳水化合物结合区域的氨基酸序列之间存在良好的相关性。
