CARBOHYDRATE BINDING PROTEIN OF B. JAPONICUM

日本大戟的碳水化合物结合蛋白

• 批准号: 3304570
• 负责人: SIUCHEONG HO
• 金额: $ 12.72万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1994-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3304570
• 关键词: Rhizobiaceae; agglutination reaction; agglutinins; bacterial genetics; bacterial proteins; cell adhesion; gene complementation; genetic manipulation; glycoprotein structure; immunoglobulins; laboratory rabbit; lectin; molecular cloning; mutant; nucleic acid sequence; protein purification; protein sequence; protein structure function; soybeans; tissue /cell culture

This project proposes to study the structure and activities of BJ38, a lectin purified from extracts of Bradyrhizobium japonicum. This protein binds to glycoconjugates containing lactose (Lac) or galactose (Gal). Derivatives of Gal at C-2, galactosamine and N-acetyl-D-galactosamine bound with much lower affinity. This specificity correlated with the behavior of intact Rhizobium in four carbohydrate-specific binding assays: (a) adsorption to Lac-Sepharose beads; (b) heterotypic binding to cultured soybean (SB-1) cells; (c) heterotypic adhesion to soybean roots; and (d) homotypic autoagglutination. These observations indicate the possibility that all of these binding activities were mediated by the same component(s) and the BJ38 is a key candidate for the carbohydrate-specific binding. The specific objectives of the proposed research are: [a] to demonstrate and quantitate the binding of purified BJ38 to soybean cells and to B. japonicum cells; [b] to test the possibility that soluble BJ38 bound to soybean cells will block adhesion of B. japonicum and that BJ38 bound to the bacteria will block autoagglutination; [c] to test whether univalent Fab fragments of antibodies directed against BJ38 will inhibit heterotypic binding to soybean cells and homotypic autoagglutination; [d] to analyze the expression and localization of BJ38 in B. japonicum cells; [e] to carry out molecular cloning of BJ38; [f] to determine the partial amino acid sequence of BJ38 at the protein level and to derive the complete amino acid sequence from the DNA sequence; [g] to test whether carbohydrate-specific binding can be restored in mutant B. japonicum by complementation with the BJ38 gene.

本项目旨在研究 BJ38 的结构和活动，BJ38 是一种 从日本慢生根瘤菌提取物中纯化的凝集素。 这种蛋白质 与含有乳糖 (Lac) 或半乳糖 (Gal) 的糖复合物结合。 Gal 在 C-2 处的衍生物、半乳糖胺和 N-乙酰基-D-半乳糖胺结合 亲和力要低得多。 这种特殊性与行为相关 四种碳水化合物特异性结合测定中的完整根瘤菌：(a) Lac-Sepharose 珠的吸附； (b) 与培养物的异型结合 大豆（SB-1）细胞； (c) 对大豆根部的异型粘附；和（四） 同型自身凝集。 这些观察结果表明了这种可能性 所有这些结合活性都是由相同的成分介导的 BJ38 是碳水化合物特异性结合的关键候选者。 拟议研究的具体目标是： [a] 证明并定量纯化的 BJ38 与大豆的结合 细胞和日本芽孢杆菌细胞； [b] 测试可溶性 BJ38 与大豆细胞结合的可能性 阻断 B. japonicum 的粘附，并且 BJ38 与细菌结合将 阻断自身凝集； [c] 测试抗体的单价 Fab 片段是否针对 BJ38 将抑制与大豆细胞的异型结合和同型结合 自身凝集； [d]分析BJ38在日本芽孢杆菌中的表达和定位 细胞； [e]进行BJ38的分子克隆； [f]确定BJ38蛋白处的部分氨基酸序列 水平并从DNA序列推导出完整的氨基酸序列； [g]测试碳水化合物特异性结合是否可以在突变体中恢复 B. japonicum 通过与 BJ38 基因互补。

项目成果

Carbohydrate binding activities of Bradyrhizobium japonicum: IV. Effect of lactose and flavones on the expression of the lectin, BJ38.

日本慢生根瘤菌的碳水化合物结合活性：IV。

• DOI: 10.1007/bf00731210
• 发表时间: 1994
• 期刊: Glycoconjugate journal
• 影响因子: 3
• 作者: Loh,JT;Ho,SC;Wang,JL;Schindler,M
• 通讯作者: Schindler,M

Carbohydrate binding activities of Bradyrhizobium japonicum: unipolar localization of the lectin BJ38 on the bacterial cell surface.

日本慢生根瘤菌的碳水化合物结合活性：凝集素 BJ38 在细菌细胞表面的单极定位。

• DOI: 10.1073/pnas.90.7.3033
• 发表时间: 1993
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Loh,JT;Ho,SC;deFeijter,AW;Wang,JL;Schindler,M
• 通讯作者: Schindler,M

Carbohydrate binding activities of Bradyrhizobium japonicum. III. Lectin expression, bacterial binding, and nodulation efficiency.

日本慢生根瘤菌的碳水化合物结合活性。

• DOI: 10.1046/j.1365-313x.1994.5060873.x
• 发表时间: 1994
• 期刊: The Plant journal : for cell and molecular biology
• 作者: Ho,SC;Wang,JL;Schindler,M;Loh,JT
• 通讯作者: Loh,JT