一种来自澳大利亚颅海绵的正常粘蛋白结合凝集素。

A normal mucin-binding lectin from the sponge Craniella australiensis.

作者信息

Xiong Chuannan, Li Wei, Liu Han, Zhang Wei, Dou Jiangli, Bai Xuefang, Du Yuguang, Ma Xiaojun

机构信息

Dalian Institute of Chemical Physics, Chinese Academy of Science, Liaoning Dalian 116023, PR China.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2006 May;143(1):9-16. doi: 10.1016/j.cbpc.2005.11.008. Epub 2006 Feb 13.

DOI:10.1016/j.cbpc.2005.11.008

PMID:16473561

Abstract

A lectin, Craniella australiensis (CAL), was isolated from sponge C. australiensis by ion-exchange on DEAE-Sephacel and purified by gel filtration on Sephadex G-150 and HPLC on DEAE-5PW. The purified lectin was a trimeric protein as revealed by SDS-PAGE and MALDI-TOF analysis. SDS-PAGE showed that the CAL protein had a molecular mass of 54 kDa, and consisted of three 18 kDa subunits. Gel filtration of purified lectin on Sephadex G-200 indicates that it exists as a 54 kDa protein in its native state. The amino acid composition was rich in Thr and Glx. CAL was found to agglutinate native and trypsinized human A, B erythrocytes, and agglutinate native erythrocytes of mouse, sheep, rabbit and chicken, and trypsinized erythrocytes of sheep and rabbit. The hemagglutination activity was inhibited by glycoproteins such as PSM and asialo-PSM, but not by any of the monosaccharides tested. The activity was stable between 20 and 70 degrees C. Significant CAL activity was observed between pH 5 and 8. The lectin reaction is independent of the presence of divalent cations Ca2+ and Mg2+. The sequence of N-terminal residues of CAL was determined as TSSCQSIVVE. The lectin showed a potent mitogenic response towards BALB/c splenocytes.

摘要

从澳大利亚海绵（Craniella australiensis）中通过DEAE - Sephacel离子交换分离出一种凝集素，即澳大利亚海绵凝集素（CAL），并通过Sephadex G - 150凝胶过滤和DEAE - 5PW高效液相色谱法进行纯化。SDS - PAGE和基质辅助激光解吸电离飞行时间（MALDI - TOF）分析表明，纯化后的凝集素是一种三聚体蛋白。SDS - PAGE显示CAL蛋白的分子量为54 kDa，由三个18 kDa的亚基组成。在Sephadex G - 200上对纯化的凝集素进行凝胶过滤表明，其天然状态下以54 kDa的蛋白形式存在。氨基酸组成富含苏氨酸（Thr）和谷氨酸（Glx）。发现CAL能凝集天然的和经胰蛋白酶处理的人A、B型红细胞，以及小鼠、绵羊、兔子和鸡的天然红细胞，还有绵羊和兔子经胰蛋白酶处理的红细胞。血凝活性受到糖蛋白如PSM和去唾液酸PSM的抑制，但不受所测试的任何单糖的抑制。该活性在20至70摄氏度之间稳定。在pH 5至8之间观察到显著的CAL活性。凝集素反应与二价阳离子Ca2 +和Mg2 +的存在无关。CAL的N端残基序列确定为TSSCQSIVVE。该凝集素对BALB/c脾细胞显示出强烈的促有丝分裂反应。