Calcium binding activity of marine invertebrate lectins and its role in biomineralization

海洋无脊椎动物凝集素的钙结合活性及其在生物矿化中的作用

• 批准号: 03660217
• 负责人: MURAMOTO Koji
• 金额: $ 1.34万
• 依托单位: TOHOKU UNIVERSITY (1992)Kitasato University (1991)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03660217/
• 关键词: Lectin; Barnacle; Invertebrate; Biomineralization; Calcium; Shell; Agglutinin; 殼

1. An enzyme-linked immunosorbent assay for the multiple lectins (BRA-2,BRA-3) of the acorn barnacle Megabalanus rosa was developed. The assay was used to study the multiple lectin levels in the coelomic fluid and various tissue homogenates of M. rosa. The concentrations of the multiple lectins ranged from 0.13 to 2.2mg/ml for BRA-2 and from 0.07 to 0.16 mg/ml for BRA-3, respectively. The protein concentration of the coelomic fluid ranged from 1.4 to 4.8mg/ml. The multiple lectins in the coelomic fluid were also identified as the major proteins by two-dimensional gel electrophoresis.2. The interaction between the multiple lectins and calcium ion was studied by ultraviolet difference spectroscopy. The association constants for BRA-2 and BRA-3 were 1.4X10^4 M^<-1> and 2.2X10^4 M^<-1> at pH8.0, respectively. The multiple lectins inhibited the crystal growth of supersaturated calcium carbonate solution at lectin concentration of 0.1mg/ml. Destruction of the carbohydrate-recognition domains of the lectins decreased the inhibitory activities on the crystal growth. Incorporation of BRAs into the crystal was demonstrated using FITC labeled lectins.

1.建立了一种测定红巨栎藤壶（Megabalanus rosa）多种凝集素（BRA-2、BRA-3）的酶联免疫吸附测定方法。该方法用于研究M.罗莎BRA-2和BRA-3的复合凝集素浓度分别为0.13 ~ 2.2mg/ml和0.07 ~ 0.16mg/ml。体腔液蛋白质浓度为1.4 ~ 4.8mg/ml。双向凝胶电泳结果表明，体腔液中的多种凝集素也是主要的蛋白质.用紫外差光谱法研究了多种凝集素与钙离子的相互作用。BRA-2和BRA-3在pH8.0时的结合常数分别为1.4 × 10^4M ^<-1>和2.2 × 10^4M ^<-1>。当凝集素浓度为0.1mg/ml时，复合凝集素对过饱和碳酸钙溶液中的晶体生长有抑制作用。糖识别结构域的破坏降低了凝集素对晶体生长的抑制活性。使用FITC标记的凝集素证明了布拉斯向晶体中的掺入。

项目成果

K.MURAMOTO: "Photoaffinity labeling of an acron barnacle lectin with a Photoactivatable fluorescent reagent derivative of D-galactosamine." Dev. Comp. Immunol.16. 1-8 (1992)

K.MURAMOTO：“用 D-半乳糖胺的光活化荧光试剂衍生物对 acron 藤壶凝集素进行光亲和标记。”

K.Muramoto: "Geographical variation of the multiple lectins in the acorn barnacle,Megabalanus rosa." Nippon Suisan Gakkaishi.

K.Muramoto：“橡子藤壶（Megabalanus rosa）中多种凝集素的地理变异。”

K.MURAMOTO: "Seasonal changes in the multiple lectin compositions of the acron barnacle Megabalanus rosa as related to ovarian development." Comp. Biochem. Physiol.98B. 603-607 (1991)

K.MURAMOTO：“藤壶Megabalanus rosa 的多种凝集素成分的季节性变化与卵巢发育有关。”

K.Muramoto: "Analysis of gamma-carboxy-glutamic acid in marine invetebrate shell matrices." Anal.Sci.7 Suppl. 939-940 (1992)

K.Muramoto：“海洋无脊椎动物壳基质中 γ-羧基-谷氨酸的分析。”

Muramoto, K. and Kamiya, H.: "The amino-acid sequence of a lectin from conger eel, Conger myriaster, skin mucus." Biochim Biophys. Acta. 1116. 129-136 (1992)

Muramoto, K. 和 Kamiya, H.：“来自海鳗、海鳗、皮肤粘液的凝集素的氨基酸序列。”