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Screening, structure and usefulness of marine bioactive proteins

海洋生物活性蛋白的筛选、结构和用途

基本信息

批准号：
08306011
负责人：
SHIOMI Kazuo
金额：
$ 5.76万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08306011/
关键词：
Marine bioactive proteins Peptide toxins Protease inhibitors Lectins Mitogens Antibacterial and antitumor proteins Amino acid sequence Gene

项目摘要

1. Two Na^+ channel toxins, one K^+ channel toxin, four novel peptide toxins and eight Kunitz-type protease inhibitors were isolated from bodies or acrorhagi of several species of sea anemones and completely sequenced.2. Acorn barnacle Megabalanus rosa lectins, BRA2 and BRA3, were successfully expressed in Escherichia coli. The sugara-binding specificity of recombinant BRA3 was the same as that of native BRA3, while recombinant BRA2 showed different sugar-binding specificity from native BRA2 probably due to the absence of carbohydrate chain in the former.3. Two rhamnose-binding lectins (STLs) were isolated from steelhead trout Oncorhynchus mykiss eggs. Sequence determinations revealed that STLs belong to a new family of animal lectins.4. A mitogenic lectin (ESA-2) recognizing specifically high mannnose type N-glycans was isolated from the red alga Euchemuaserra. It contained several repeated sequences in the molecule, showing high sequence similarity with some bacterial hemagglutinins. Furthermore, it inhibited the growth of cultured human tumor cells.5. An aplysianin-P (antibacterial glycoprotein from the purple fluid of sea hare Aplysia kurodai) cDNA clone was not obtained by either RT-PCR method or screening of cDNA libraries from purple gland, albumen gland and midgut. This may suggest that the gene of aplysianin-P is not expressed in the tissues examined or that the previouly determined amino acid sequence of aplysianin-P is incorrect.6. Hemolytic and lectin activities of the eel skin mucus decreased with the rise of antibody activity in the blood plasma. Purified skin mucus lectin was found to have both opsonic and chemotactic activities against own neutrophiles. These results mean the cooperative action of surface and inner defense mechanisms. High disease susceptibility of larvae was seen in abalone born from a mother of low lectin activity, suggesting that the lectin activity is one of the characters of disease resistance in abalone.

1.从几种海葵的体或囊体中分离到两种Na^+通道毒素、一种K^+通道毒素、四种新的肽类毒素和八种Kunitz型蛋白酶抑制剂，并进行了全序列测定.在大肠杆菌中成功表达了橡实藤壶Megabalanus rosa凝集素BRA 2和BRA 3。重组BRA 3的糖结合特异性与天然BRA 3相同，而重组BRA 2的糖结合特异性与天然BRA 2不同，可能是由于重组BRA 2中缺少糖链.从虹鳟（Oncorhynchus mykiss）卵中分离到两种鼠李糖结合凝集素（STL）。序列测定表明，STLs属于一个新的动物凝集素家族.从红毛桉中分离到一种特异识别高甘露糖型N-聚糖的促有丝分裂凝集素（ESA-2）。它在分子中含有几个重复序列，显示出与某些细菌血凝素的高度序列相似性。此外，它还能抑制体外培养的人肿瘤细胞的生长.通过RT-PCR方法和从紫腺、蛋白腺和中肠cDNA文库中筛选，均未获得一个黑腹海兔紫液抗菌糖蛋白（Aaprosianin-P）的cDNA克隆。这可能表明，在所检测的组织中，花生素-P的基因不表达，或者先前测定的花生素-P的氨基酸序列是不正确的。鳗鱼皮肤粘液的溶血活性和凝集素活性随血浆抗体活性的升高而降低。纯化的皮肤粘液凝集素被发现具有调理和趋化活性，对自己的嗜中性粒细胞。这些结果表明，表面和内部防御机制的协同作用。低凝集素活性的母鲍所产幼鲍对疾病的易感性高，表明凝集素活性是鲍的抗病性特征之一。