Lipopolysaccharides in Pseudomonas aeruginosa : Structure and biological activity of D-rhamnan-linked lipopolysaccharides

铜绿假单胞菌中的脂多糖：D-鼠李聚糖连接脂多糖的结构和生物活性

• 批准号: 62470146
• 负责人: ARAKI Yoshio
• 金额: $ 2.94万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62470146/
• 关键词: D-Rhamnan; Common antigen; Lipopolysaccharide; Pseudomonas aeruginosa; Monoclonal antibody; D-rhamnan-linked lipopolysaccharide; O-多糖; モノクロナル抗体; グラム陰性菌; Oー抗原多糖; Dーラムナン; 単クローン抗体; マクレオチド糖

Recently, a monoclonal antibody E87 has been developed by Sawada et al. From the wide cross-reactivity againsy P. aeruginosa isolates, this antibody seems to be useful in immunotherapy for P. aeruginosa infection. In this project the occurrence and structure of E87 antigen were studied, and the following results are obtained.(1) Rhamnose-rich polysaccharides were isolated from the O-polysaccharide fractions of three P. aeruginosa strains, IFO 3080 (Homma serotype M), IID 1020 (serotype G), and T424 (serotype G), after selective degradation degradation of the respective major O-antigens. On the basis of their chemical composition, measurement of ^1H - and ^<13>C-NMR spectra, and determination of optical rotatory, it is concluded that these polysaccharides have the same D-rhamnan structure, -3)-D-Rha(alpha1->2)-D-Rha(alpha1->3)-D-Rha(alpha1->, just like D-rhamnan from the strain IID 1008.(2) In competitive ELISA, the above rhamnose-rich polysaccharide preparations as well as the D-rhamnan of strain IID 1008 inhibited, in the same extent, the binding of monoclonal antibody E87 to P. aeruginosa PAC 1 cells. Thus, the isolated polysaccharides are immunologically indistinguishable from the D-rhamnan.(3) The enzyme assay system for GDP-D-rhamnose synthetase was developed, and it was shown that this enzyme occurs in nine (A, D, F, G, H, I, K, L, and M) of tested thirteen serotypes of P. aeruginosa. This enzyme distribution was closely correlated to the binding spectrum of monoclonal antibody E87 to these cells.The above results strongly supports a possibility that E87 antigen is D-rhamnan- linked lipopolysaccharides. From their wide occurrence, the D-rhamnan-linked lipo polysaccharide may be a common antigen in P. aeruginosa.

最近，Sawada等人开发了一种单克隆抗体E87。从对铜绿假单胞菌分离物的广泛交叉反应性来看，该抗体似乎可用于铜绿假单胞菌感染的免疫治疗。本课题对E87抗原的发生和结构进行了研究，得到如下结果：(1)从3株P. aeruginosa菌株IFO 3080 （Homma血清型M）、IID 1020（血清型G）和T424（血清型G）的o -多糖中选择性降解各主要o -抗原，分离得到富鼠李糖多糖。根据它们的化学成分、^1H -和^<13>C-NMR谱的测量以及旋光度的测定，得出这些多糖具有相同的D-rhamnan结构，-3)- d -rha (alpha1->2)- d -rha (alpha1->3)- d -rha (alpha1->3)- d -rha (alpha1->)，与菌株iid1008中的D-rhamnan相同。(2)在竞争性ELISA中，上述富鼠李糖多糖制剂和菌株iid1008的d -鼠李糖均在相同程度上抑制了单克隆抗体E87与铜绿假单胞菌PAC 1细胞的结合。因此，分离的多糖在免疫学上与d -鼠李糖难以区分。(3)建立了gdp -D-鼠李糖合成酶酶分析系统，结果表明该酶存在于13种铜绿假单胞菌血清型中的A、D、F、G、H、I、K、L、M 9种酶中。该酶的分布与单克隆抗体E87与这些细胞的结合谱密切相关。上述结果有力地支持了E87抗原为d -鼠李糖链脂多糖的可能性。从它们的广泛存在来看，d -鼠李糖链脂多糖可能是铜绿假单胞菌的共同抗原。

项目成果

N. Murazumi, K. Kumita, Y. Araki, & E. Ito: "Partial purification and properties of UDP-N-acetylmannos-amine:N-acetylglucosaminyl pyrophosphorylundecaprenol N-acetylmannosaminyltransferase from Bacillus subtilis" J. Biochem. 104, 980-984, 1988.

N. Murazumi、K. Kumita、Y. Araki、

S. Kaya, Y. Araki, & E. Ito: "The structure of the O-polysaccharide chain of lipopolysaccharide from Pseudomonas aeruginosa IID 1012 (ATCC 27588)" J. Biochem. 105, 29-34, 1989.

S.卡亚，Y.荒木，

H.Iwasaki,Y.Araki,S.Kaya,E.Ito: "Structural studies on the amino-acid-linked teichuronic acid of Bacillus subtilis AHU 1219 cell walls" European Journal of Biochemestry. 178. 643-648 (1989)

H.Iwasaki、Y.Araki、S.Kaya、E.Ito：“枯草芽孢杆菌 AHU 1219 细胞壁的氨基酸连接的 Teichuronic 酸的结构研究”欧洲生物化学杂志。

Shin-ichi,Yokota: European Journal of Biochemistry. 167. 203-209 (1987)

横田信一：欧洲生物化学杂志。

Yoshio Araki: Biochemistry.

荒木芳雄：生物化学。