Research for a basic anti-microbial recognition system of the animal using silkworm, Bombyx mori

家蚕动物基本抗菌识别系统的研究

基本信息

批准号：
10680601
负责人：
SATO Ryoichi
金额：
$ 1.98万
依托单位：
Tokyo University of Agriculture & Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680601/
关键词：
C-type lectin BmLBP Bombyx mori recognition protein Bombyx mori

项目摘要

Affinity purification was conducted with microbial cells to obtain anti-microbial recognition proteins from the larval hemolymph of the silkworm. Bombyx mori.A 43 kDa lipopolysaccharide-binding protein was purified with Esherichia coli cells and named BmLBP. BmLBP bound to many kinds of gram-negative bacteria by recognition of lipid A and helped nodule formation of the blood cells. From the result of cDNA cloning and sequencing it was shown that BmLBP is a novel member of the C-type lectin superfamily with a unique structural feature that consists of two different carbohydrate-recognition domains in tandem.Two proteins of 42 kDa were purified respectively with Saccharomyces cervisiae cells and Micrococcus luteus cells. Both bound bacteria by recognizing carbohydrate of the bacterial cell wall. Also, they needed calcium in binding. From these characters two protein were assumed to be C-type lectins.Antisera were constructed against these three binding proteins and closs-reactivity was tested each other. As a result, two protein obtained with Saccharomyces cervisiae cells and Micrococcus luteus cells were shown to be selologically the same and BmLBP was shown to be different from these two proteins. From these results it was supposed that only two kinds of lectin is sufficient to recognize fairly broad range of microorganisms.

利用微生物细胞进行亲和纯化，从家蚕幼虫血淋巴中获得抗微生物识别蛋白。家蚕(Bombyx mori)。用大肠杆菌细胞纯化了43 kDa的脂多糖结合蛋白，并将其命名为BmLBP。 BmLBP通过识别脂质A与多种革兰氏阴性菌结合，并帮助血细胞结节的形成。 cDNA克隆和测序结果表明，BmLBP是C型凝集素超家族的新成员，具有独特的结构特征，由两个不同的碳水化合物识别域串联组成。分别纯化了两种42 kDa的蛋白质。酿酒酵母细胞和藤黄微球菌细胞。两者都通过识别细菌细胞壁的碳水化合物来结合细菌。此外，它们还需要钙来结合。根据这些特征，假定两种蛋白是C型凝集素。针对这三种结合蛋白构建了抗血清，并测试了彼此的紧密反应性。结果，用酿酒酵母细胞和藤黄微球菌细胞获得的两种蛋白质在血清学上是相同的，而BmLBP与这两种蛋白质不同。根据这些结果，推测仅两种凝集素就足以识别相当广泛的微生物。