Siderophore-Mediated Iron-Uptake System in Vibrio parahaemolyticus and Its Relevance to Pathogenesis

副溶血弧菌铁载体介导的铁摄取系统及其与发病机制的相关性

• 批准号: 05670257
• 负责人: YAMAMOTO Shigeo
• 金额: $ 1.34万
• 依托单位: Okayama University, Faculty of Pharmaceutical Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670257/
• 关键词: Vibrio parahaemolyticus; Iron assimilation; Siderophore; Iron source; Outer membrane receptor; Transferrin utilization; Hemin utilization; Pathogenesis; ヘミン利用性; ヘミンレセプター; 病原因子; 外膜蛋白

1.The function of vibrioferrin (VE) as a siderophore in iron-starved Vibrio parahaemolyticus was demonstrated by uptake of [55Fe] ferric VF displaying kinetics typical of a protein receptor-mediated process. The iron uptake mediated by VF was blocked by uncouplers and ATPase inhibitors. The EDDA-plate assays showed that VF was inactive to other Vibrio Species except for V.alginolyticus, which was found later to synthesize VF.2.The amounts of VF secreted to the culture supernatants were quantified by HPLC for 32 strains of V.parahaemolyticus isolated from different sources. The clinical isolates (n=8,33.6muM) produced VF 10-fold over the environmental isolates (n=22,3.9muM), indicating that the producibility of VF is advantageous of surviving and proliferating in human hosts.3.The ability of VF to assimilate iron for growth from 30% iron-saturated human transferrin was demonstrated by comparison of growth rate between strain AQ 3354 and its spontaneous mutant defective in VF synthesis. … More This suggests that the organism may utilize such a source of host iron through the action of VF during in vivo survival and proliferation.4.Under iron-restricted conditions, V.parahaemolyticus expressed two major outer membrane proteins of 78 and 83 kDa. Autoradiographic analysis of outer membrane preparations previously incubated with [55Fe] -VF revealed a single radiolabeled band, in which the 78-kDa protein was detected predominantly. The 78-kDa protein was cleaved by the treatment of whole cells with proteinase K,indicating its cell surface-exposed location. These results suggest that the Fe-VF binding protein of 78 kDa may function as the receptor for Fe-VF.Immunoblot analysis using the antiserum raised against the purified 78-kDa protein indicated that the molecular mass and antigenic properties of the protein were highly conserved among this species.5.Several clinical isolates were examined for their ability to utilize either hemin or hemoglobin as a sole source of iron. Both compounds appeared to be equally good iron sources. The hemin-agarose batch affinity method allowed us to identify the 83-kDa protein as the hemin-binding protein.6.Possession of two iron accquisition systems may be important at different stages or sites of infection, or as insurance against a mutational loss of one of them. The preparation of mutants defective of either or both of these receptor proteins is under progress to evaluate the contribution of these iron uptake systems to the physiology and infectious process of V.parahaemolyticus. Less

1.铁蛋白（VE）作为铁载体在铁饥饿的副溶血性弧菌的功能被证明通过摄取[55 Fe]铁VF显示动力学典型的蛋白质受体介导的过程。VF介导的铁摄取可被解偶联剂和ATP酶抑制剂阻断。EDDA平板法显示，VF对其他弧菌无活性，但对溶藻弧菌（V.alginolyticus）无活性，后者是后来发现的合成VF的弧菌。临床分离株（n= 8，33.6 μ M）产生的VF是环境菌株的10倍（n= 22，3.9 μ M），表明VF的产生有利于其在人体内的存活和增殖。通过比较菌株AQ 3354和其VF合成缺陷的自发突变体之间的生长速率，证明了饱和的人转铁蛋白。 ...更多信息 这表明，在体内生存和增殖过程中，该菌可能通过VF的作用来利用宿主铁源。4.在铁限制条件下，副溶血性弧菌表达两种主要的外膜蛋白，分子量分别为78和83 kDa。预先与[55 Fe] -VF孵育的外膜制备物的放射自显影分析显示单一放射性标记带，其中主要检测到78 kDa蛋白。78 kDa的蛋白质被裂解的处理与蛋白酶K的整个细胞，表明其细胞表面暴露的位置。这些结果表明，78 kDa的Fe-VF结合蛋白可能是Fe-VF的受体。免疫印迹分析表明，该蛋白的分子量和抗原特性在该物种中是高度保守的。这两种化合物似乎是同样好的铁源。氯化血红素-琼脂糖批次亲和法使我们能够确定83-kDa的蛋白质作为氯化血红素结合蛋白。6.拥有两个铁积累系统可能是重要的，在不同的阶段或感染的网站，或作为保险对突变的损失之一。这些受体蛋白的突变体缺陷的制备正在进行中，以评估这些铁吸收系统的生理和感染过程的副溶血性弧菌的贡献。少

项目成果

Shigeo Yamamoto: "Utilization of hemin and hemoglobin as iron sources by Vibrio parahaemolyticus and identification of an iron-repressible hemin-binding protein" FEMS Microbiology Letters. (in press). (1995)

Shigeo Yamamoto：“副溶血弧菌利用血红素和血红蛋白作为铁源以及铁抑制血红素结合蛋白的鉴定”FEMS 微生物学快报。

S.Yamamoto: "Utilization of Hemin and Hemoglobin as an Iron Sources by Vibrio parahaemolyticus and Identification of an Iron-Repressible Hemin-Binding Protein" FEMS Microbiology Letters. (in press). (1995)

S.Yamamoto：“副溶血弧菌利用血红素和血红蛋白作为铁源以及铁抑制血红素结合蛋白的鉴定”FEMS 微生物学快报。

Shigeo Yamamoto: "Structure and iron transport activity of vibrioferrin,a new siderophore of Vibrio parahaemolyticus" Journal of Biochemistry. 115. 868-874 (1994)

Shigeo Yamamoto：“副溶血弧菌的新铁载体弧铁蛋白的结构和铁转运活性”生物化学杂志。

Shigeo Yamamoto: "Demonstration of a ferric vibrioferrin-binding protein in the outer membrane of Vibrio parahaemolyticus" Microbiology and Immunology. (Submitted for publication).

Shigeo Yamamoto：“副溶血弧菌外膜中铁弧菌铁蛋白结合蛋白的演示”微生物学和免疫学。

S.Yamamoto: "Siderophore-Mediated Utilization of Iron Bound to Transferrin by Vibrio parahaemolyticus" Microbiology and Immunology. 38. 687-693 (1994)

S.Yamamoto：“副溶血弧菌介导的铁载体介导的与转铁蛋白结合的利用”微生物学和免疫学。