Adhesion Molecule of Borrelia burgdorferi ; Its Function and Application

伯氏疏螺旋体的粘附分子；

• 批准号: 09670294
• 负责人: MASUZAWA Toshiyuki
• 金额: $ 1.41万
• 依托单位: University of Shizuoka
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670294/
• 关键词: Lyme disease; Borrelia; Virulence factor; Adhesion molecule; Glycosphingolipid; Galactosylceramide; Borrelia burgdorferi; Glyceradehyde-3-phosphate dehydrogenase; カラクトシルセラミド; グリセルアルデヒド-3ーリン酸脱水素酵素

The binding activities of Borrelia burgdorferi to glycosphingolipid which are present in various types of cells were examined. B.burgdorferi bound specifically to galactosylceramide (GalCer) and glucosylceramide (GicCer), but not to other glycosphingolipids. The binding specificity of B.burgdorferi to various glycosphingolipid suggested that the binding receptor in the species is ceramide monohexoside. The necessity of the sugar and N-acyl moieties in galactosylceramide for the binding of Borre1ia was shown using chemically modified GalCer. The binding activities of the low-passage virulent strains to GalCer on TLC plates and to CHO-K1 cells in vitro were threefold higher than those of high-passage avirulent strains. Binding was no affected by pretreatment of Borrelia with monospecific anti-outer surface protein C (OspC) serum. These results indicated that the binding of Borrelia to glycosphingolipid expressed cell surface play an essential role in infection of mammalian hosts, but that OspC is not associated with binding. Two proteins, 37kDa and 67kDa, were involved in binding of B.burgdorferi to galactosylceramide as shown by blotting assay using biotinylated GalCer as a probe. 37kDa and 67kDa proteins reactive were determined as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and homologue of heat shock protein 62, respectively. Recombinant GAPDH expressed by E.coli were bound with GalCer on TLC plate. The finding obtained in this study indicated that GAPDH is one of adhesion factor of B.burgdorferi.

检测了伯氏疏螺旋体与存在于各种类型细胞中的鞘糖脂的结合活性。B.burgdorferi特异性结合半乳糖神经酰胺（GalCer）和葡萄糖神经酰胺（GicCer），但不结合其他鞘糖脂。B.burgdorferi与多种鞘糖脂结合的特异性表明，该种结合受体为神经酰胺单己糖苷。使用化学修饰的GalCer显示了半乳糖神经酰胺中的糖和N-酰基部分用于Borre 1 ia结合的必要性。低代毒力菌株与薄层板上的GalCer结合活性和与CHO-K1细胞的体外结合活性均比高代无毒力菌株高3倍。结合不受用单特异性抗外表面蛋白C（OspC）血清预处理疏螺旋体的影响。这些结果表明疏螺旋体与表达鞘糖脂的细胞表面的结合在哺乳动物宿主的感染中起重要作用，但OspC与结合无关。以生物素标记的GalCer为探针进行的印迹分析表明，37 kDa和67 kDa两种蛋白参与了伯氏B菌与半乳糖神经酰胺的结合。37 kDa和67 kDa蛋白分别被确定为甘油醛-3-磷酸脱氢酶（GAPDH）和热休克蛋白62的同源物。将大肠杆菌表达的重组GAPDH与TLC板上的GalCer结合。本研究结果表明GAPDH是伯氏疏螺旋体的粘附因子之一。

项目成果

Wang, J.et al.: "Sequence analysis of OspA and OspB genes of Borrelia garinii isolated from patient in Japan and its evolutionary implications." FEMS Microbiol.Lett.154. 317-375 (1997)

Wang, J.等人：“从日本患者体内分离出的加氏疏螺旋体 OspA 和 OspB 基因的序列分析及其进化意义。”

Kazuhide Kaneda, Toshiyuki Masuzawa, Kitsuzo Yasugami, Takashi Suzuki, Yasuo Suzuki, and Yasutake Yanagihara: "Identification of glycosphingolipid-binding protein of Borrelia burgdorferi sensu lato." Infect.Immun.65. 3180-3185 (1997)

Kazuhide Kaneda、Toshiyuki Masuzawa、Kitsuzo Yasugami、Takashi Suzuki、Yasuo Suzuki 和 Yasutake Yanagihara：“伯氏疏螺旋体鞘糖脂结合蛋白的鉴定。”

矢島あゆみ他: "刺し口の周囲にしびれ感を伴ったオーストラリア毒マダニ刺咬症の1例"皮膚. 40. 399-402 (1998)

Ayumi Yajima 等人：“澳大利亚有毒蜱虫叮咬并伴有穿刺部位周围麻木的案例”皮肤。 40. 399-402 (1998)

Toshiyuki Masuzawa, Takako Fukui, Masaki Miyake, Hee-Bok Oh, Min-Kee Cho, Woo-Hyun Chang, Yasuyuki Imai, and Yasutake Yanagihara, Y: "Determination of Borrelia afzelii-related group isolated from Ixodes nipponensis in Korea as Borrelia valaisiana."Int. J.

Toshiyuki Masuzawa、Takako Fukui、Masaki Miyake、Hee-Bok Oh、Min-Kee Cho、Woo-Hyun Chang、Yasuyuki Imai 和 Yasutake Yanagihara，Y：“将韩国从日本硬蜱中分离出的阿氏疏螺旋体相关群确定为瓦莱疏螺旋体

Jianhui Wang, Toshiyuki Masuzawa, and Yasutake Yanagihara.: "Sequence analysis of OspA and OspB genes of Borrelia garinii isolated from patient in Japan and its evolutionary implications." FEMS Microbiol.Lett.154. 371-375 (1997)

Kenhui Wang、Toshiyuki Masuzawa 和 Yasutake Yanagihara：“从日本患者体内分离出的加氏疏螺旋体 OspA 和 OspB 基因的序列分析及其进化意义。”