Amino Acid Sequence and Carbohydrate-binding Analysis of the N-acetyl-D-galactosamine-specific C-Type Lectin, CEL-I, from the Holothuroidea, Cucumaria echinata
Amino Acid Sequence and Carbohydrate-binding Analysis of the N-acetyl-D-galactosamine-specific C-Type Lectin, CEL-I, from the Holothuroidea, Cucumaria echinata
复制标题
海参科 N-乙酰基-D-半乳糖胺特异性 C 型凝集素 CEL-I 的氨基酸序列和碳水化合物结合分析
DOI：
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
H. Aoyagi
中科院分区：
文献类型：
--
作者：
T. Hatakeyama;Noriaki Matsuo;Kouhei Shiba;S. Nishinohara;N. Yamasaki;H. Sugawara;H. Aoyagi
CEL-I是从海参（Cucumaria echinata）中分离得到的一种钙依赖性凝集素。这种蛋白质由两个相同的亚基组成，由一个二硫键连接。通过对S-吡啶基乙基化的CEL-I的蛋白水解片段化产生的肽进行测序来确定CEL-I的完整氨基酸序列。CEL-I的亚基由140个氨基酸残基组成。两个链内（Cys 3-Cys 14和Cys 31-Cys 135）和一个链间（Cys 36）二硫键也被确定从分析的含胱氨酸的肽从完整的蛋白质。CEL-I的序列与其他C型凝集素的序列相似性较低，而C端区域，包括推定的Ca 2+和碳水化合物结合位点，相对保守。当碳水化合物结合活性进行了检查，通过固相微孔板测定，CEL-I显示出更高的亲和力N-乙酰基-D-半乳糖胺比其他半乳糖相关的碳水化合物。测定了CEL-I与对硝基苯基N-乙酰-β-D-氨基半乳糖胺（NP-GalNAc）的结合常数为2.3×104 M ~ 1，平衡透析实验测得NP-GalNAc的最大结合数为1.6。
CEL-I is one of the Ca2+-dependent lectins that has been isolated from the sea cucumber, Cucumaria echinata. This protein is composed of two identical subunits held by a single disulfide bond. The complete amino acid sequence of CEL-I was determined by sequencing the peptides produced by proteolytic fragmentation of S-pyridylethylated CEL-I. A subunit of CEL-I is composed of 140 amino acid residues. Two intrachain (Cys3-Cys14 and Cys31-Cys135) and one interchain (Cys36) disulfide bonds were also identified from an analysis of the cystine-containing peptides obtained from the intact protein. The similarity between the sequence of CEL-I and that of other C-type lectins was low, while the C-terminal region, including the putative Ca2+ and carbohydrate-binding sites, was relatively well conserved. When the carbohydrate-binding activity was examined by a solid-phase microplate assay, CEL-I showed much higher affinity for N-acetyl-D-galactosamine than for other galactose-related carbohydrates. The association constant of CEL-I for p-nitrophenyl N-acetyl-β-D-galactosaminide (NP-GalNAc) was determined to be 2.3×104 M1, and the maximum number of bound NP-GalNAc was estimated to be 1.6 by an equilibrium dialysis experiment.
影响因子：
2.9
作者：
GILL, SC;VONHIPPEL, PH
通讯作者：
VONHIPPEL, PH
基于 GaN 量子阱的 249 nm 深紫外激光器
影响因子：
7
作者：
Shan, Maocheng;Zhang, Yi;Li, Xiaohang
通讯作者：
Li, Xiaohang