Arata Yoichiro, Tamura Mayumi, Nonaka Takamasa, Kasai Ken-ichi
Department of Biological Chemistry, Teikyo University School of Pharmaceutical Sciences, Sagamiko, Kanagawa 199-0195, Japan.
Biochem Biophys Res Commun. 2006 Nov 10;350(1):185-90. doi: 10.1016/j.bbrc.2006.09.017. Epub 2006 Sep 15.
The N-terminal lectin domain (Nh) of the tandem repeat-type nematode galectin LEC-1 has a lower affinity for sugars than the C-terminal lectin domain. To confirm that LEC-1 forms a complex with N-acetyllactosamine-containing glycoproteins, we used several mutants of LEC-1 in which a unique cysteine residue was introduced into the Nh domain and examined their binding to bovine asialofetuin with a photoactivatable sulfhydryl crosslinking reagent. A crosslinked product was formed with the Q38C mutant, strongly suggesting the low-affinity interaction of Nh with the glycoprotein could be detected with this system.
串联重复型线虫半乳糖凝集素LEC-1的N端凝集素结构域（Nh）对糖的亲和力低于C端凝集素结构域。为了证实LEC-1与含N-乙酰乳糖胺的糖蛋白形成复合物，我们使用了几种LEC-1突变体，其中在Nh结构域引入了一个独特的半胱氨酸残基，并用可光活化的巯基交联剂检测它们与牛去唾液酸胎球蛋白的结合。Q38C突变体形成了交联产物，强烈表明该系统能够检测到Nh与糖蛋白的低亲和力相互作用。
