3-Dimensional structure of staphylococcal leukocidin and γ-hemolysin

葡萄球菌杀白细胞素和 γ-溶血素的 3 维结构

• 批准号: 11694191
• 负责人: KAMIO Yoshiyuki
• 金额: $ 3.46万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694191/
• 关键词: Staphylococcus aureus; pore-forming toxin; γ-hemolysin; leukocidin; staphylococcal gemone; phage conversion; 3-dimensional structure; 2成分性毒素; 三次元構造; X線回折

Crystal Structure of LukF Delineates Conformational Changes Accompanying For-mation of a Transmembrane ChannelLukF Architecture. LukF has the shape of a prolate ellipsoid with dimensions of 72Åx34Åx25. In LukF the amino latch and the pre-stem adopt dramatically different conformations when compared to the corresponding regions of an Hla protomer. Excluding these areas, the fold of LukF is identical to the fold of an Hla protomer. The residues that comprise the amino latch (E2-K16) adopt a β-strand conformation and extend the inner β-sheet of the β-sandwich by one strand in LukF.Measured in terms of solvent accessible surface area, binding amino latch to the LukF core buries 414 Å^2. In striking contrast ot the Hla protomer, the glycine-rich region of LukF forms a three-strand antiparalleled β-sheet that packs against the inner β-sheet of the β-sandwich domain. The pre-stem β-sheet, which includes the seven disordered residues spanning strand 7" and strand 8 (Figures 1C and 10), has an … More a(β)b(β)c(coil)d(β) fold. Hydrophobic residues predominate in the pre-stem interface with the β-sandwich domain : there are 24 van der Walls interactions and the buried surface area is 852Å^2, which is close to the 670 Å^2 buried by the Hla amino latch when bound at a similar site on an adjacent protomer in the heptamer structure. Direct contact between the pre-stem and the amino latch is mediated by a cluster of hydrophobic residues that includes V13, V17, Y117, and F119. In Hla, the residues that occupy the latter two positions are predicted to lie at the interfacial region of the lipid bilayer. The juxtaposition of Y117 and F119 between the pre-stem and the amino latch makes direct "communication" between these two key regions possible.LukF Surface Properties and Phospholipid Binding Site. The surface of the pre-stem β-sheet facing the LukF core is primarily hydrophobic while the side directed toward the solvent is polar. Like Hla, the LukF rim domain contains a lot of exposed aromatic residues. Located in a cleft lined by W177 and R198 is a binding site for phospholipid head groups. In the Hla heptamer, there is a similar lipid binding site, i.e., W179 and R200 residues of Hla, and the latter residue is critical for binding of Hla to an erythrocyte membrane.LukF Monomer and Hla Protomer Comparison. Superposition of the LukF and Hla protomer structures reinforces the conclusion that the cores are very similar despite a sequence identity of only 31.7% for the mature polypeptides. The comparison also emphasizes the divergence in conformation at the amino latch and glycine-rich "stem" regions. Fitting of individual domains demonstrated that the r.m.s deviation between Cα positions for the β-sandwich and rim are 2.1 Å and 2.4 Å, respectively. To a first approximation, the β-sandwich and rim domains behave as rigid bodies that adopt different relative conformations in the monomer (LukF) and heptamer (Hla) due to small changes spread over a number of residues at the β sandwich/rim domain juncture. Recently, the 3-dimensional structure of the water-soluble LukF-PV monomer also analysed and was found to be basically similar to that of LukF.Major conformational differences between LukF and the Hla protomer structures occur in the triangle region. In contrast to the diffuse nature of the conformational differences relating the β- sandwich and rim domains, there are larger differences in main chain phi and psi angles for 4 residues in the triangle region which allow the LukF pre-stem to fold against the inner surface of the β-sandwich domain. In the Hla protomer, these main chain dihedral angles differ by>90゜ and are associated with a triangle conformation in which the polypeptide chain extends from the protomer core. The LukF pre-stem occupies approximately the same site that the amino latch of a neighboring protomer occupies in the oligomeric toxin, based on the analogy with the Hla heptamer. Less

LUKF的晶体结构描绘了变化的变化，即在LUKF中的尺寸为72Åx34Åx25的跨膜结构。 HLA原始物的折叠。采用β链构象，并将β-sandwich的故事扩展到lukf中的一个链。用术语计算，将氨基闩锁与lukf core buries buries buriesh rast rast rast rast rast LUKF的富含甘氨酸的区域形成了β-Sandwich结构域的内部表的三链抗β-片，跨越了链7”，并具有链8（图1C和10）。 ）B（线圈）D（β）折叠。七聚体结构中的相似位点。脂质双层。侧向HLA的侧面，LUKF RIM域中包含大量暴露的芳族残基。对于hlathrocyte膜的结合至关重要。LUKF单体和HLA frotomer比较，尽管在成熟多肽的置信度中也有31.7％的序列，但HLA frotomer结构加强了核心的结论。氨基闩锁和富含甘氨酸的“ g” g表明，β-sandwich和RIM的Cα位置之间的R.M.S偏差为2.1Å2.4Å，首次参与，β-Sandwich和RIM域是刚性的在E单体（LUKF）中采用不同相对构象的身体，由于β夹心/rimain联结的许多居民的少量变化而导致的体积也很小。分析到LUKF的基本伊米尔。与lukf和HLA frotomer结构之间的汇总差异发生在三角形区域中。 lukf预先茎可以折叠到hla frotomer的内表面，主链二进制角度不同于> 90，其中多肽链从frotomer核心延伸。邻近的原始寡聚毒素的氨基闩锁

项目成果

H.Katsumi, T.Tomita, J.Kaneko, and Y.Kamio: "Vitronectin and its fragments purified as serum inhibitors of Staphylococcus aureus gammahemolysin and leukocidin, and their specific binding to the Hlg2 and the LukS components of the toxins"FEBS Lett.. 460. 4

H.Katsumi、T.Tomita、J.Kaneko 和 Y.Kamio：“玻连蛋白及其片段纯化为金黄色葡萄球菌伽玛溶血素和杀白细胞素的血清抑制剂，以及它们与毒素的 Hlg2 和 LukS 成分的特异性结合”FEBS Lett

Y.Kamio: "TRENDS in PROTEIN RESEARCH"Scientific Publishers OWN. 150 (1999)

Y.Kamio：《蛋白质研究趋势》科学出版社拥有。

神尾好是: "黄色ブドウ球菌の細胞崩壊毒素の遺伝子、構造、および作用機構の解明"日本農芸化学会誌. 73(7). 703-711 (1999)

Yoshiharu Kamio：“金黄色葡萄球菌细胞毒素的基因、结构和作用机制的阐明”，日本农业化学学会杂志 73(7) (1999)。

金子淳: "黄色ブドウ球菌の二成分細胞崩壊毒素のファージ変換及び標的細胞との作用に関する研究"日本農芸化学会誌. 75巻(印刷中). (2001)

Jun Kaneko：“金黄色葡萄球菌二元溶细胞毒素的噬菌体转化及其与靶细胞的相互作用”，日本农业化学学会杂志，第 75 卷（出版中）。

D.Zou, J.Kaneko, S.Narita, and Y.Kamio: "Prophage, φPV83-pro, carrying Panton-Valentine leukocidin genes, on the Staphylococcus aureus P83 chromosome : comparative analysis of the genome structures of φPV837-pro, φPVL, φ11, and other phage"Biothechnol.Bio

D.Zou、J.Kaneko、S.Narita 和 Y.Kamio：“Prophage，φPV83-pro，在金黄色葡萄球菌 P83 染色体上携带 Panton-Valentine 杀白细胞素基因：φPV837-pro、φPVL 基因组结构的比较分析、 φ11 等噬菌体“Biothechnol.Bio