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的晶体结构描绘了伴随着跨膜细胞LukF结构形成的构象变化。LukF的形状是一个长椭圆体，尺寸为72 x34 x25。在LukF中，与Hla原聚体的相应区域相比，氨基闩锁和前茎采用显著不同的构象。排除这些区域，LukF的折叠与Hla原聚体的折叠相同。构成氨基锁的残基（E2-K16）采用β链构象，并将LukF中β夹心结构的内部β折叠延伸了一条链，根据溶剂可及表面积测量，将氨基锁结合到LukF核心上可埋414个碱基^[2]。与Hla原聚体形成鲜明对比的是，LukF的富含甘氨酸的区域形成了一个三链抗β折叠，该折叠与β夹心结构域的内部β折叠相挤压。包括跨越链7”和链8的七个无序残基的前茎β-折叠（图1C和10）具有与链7“和链8”相同的长度。 ...更多信息 a（β）B（β）c（coil）d（β）折叠。疏水残基在前茎界面与β-夹心结构域中占主导地位：有24个货车德尔壁相互作用，掩埋表面积为852平方厘米，这接近于Hla氨基闩锁在七聚体结构中相邻原聚体上类似位点结合时掩埋的670平方厘米。前茎和氨基闩锁之间的直接接触由包括V13、V17、Y117和F119的疏水残基簇介导。在Hla中，占据后两个位置的残基被预测位于脂质双层的界面区域。Y117和F119在前茎和氨基锁之间的并置使得这两个关键区域之间的直接“通信”成为可能。LukF表面性质和磷脂结合位点。面向LukF核心的预茎β-折叠的表面主要是疏水性的，而朝向溶剂的一侧是极性的。与Hla一样，LukF边缘结构域含有大量暴露的芳香残基。位于W177和R198之间的裂缝是磷脂头基的结合位点。在Hla七聚体中，存在类似的脂质结合位点，即，Hla的W179和R200残基，并且后者残基对于Hla与红细胞膜的结合是关键的。LukF和Hla原聚体结构的叠加强化了以下结论：尽管成熟多肽的序列同一性仅为31.7%，但核心非常相似。该比较还强调了在氨基闩锁和富含甘氨酸的“茎”区域的构象的分歧。单个结构域的拟合表明，β-夹心和边缘的Cα位置之间的r.m.s偏差分别为2.1 μ m和2.4 μ m。对于第一近似，β-夹心和边缘结构域表现为刚性体，其由于在β夹心/边缘结构域接合处的许多残基上的小变化而在单体（LukF）和七聚体（Hla）中采用不同的相对构象。最近，水溶性LukF-PV单体的三维结构也进行了分析，发现LukF的三维结构基本相似，LukF和Hla原聚体结构之间的主要构象差异发生在三角形区域。与β-夹心结构域和边缘结构域相关的构象差异的扩散性质相反，三角形区域中4个残基的主链phi和psi角存在较大差异，这允许LukF前茎折叠在β-夹心结构域的内表面上。在Hla原聚体中，这些主链二面角相差>90 °，并且与其中多肽链从原聚体核心延伸的三角形构象相关。基于与Hla七聚体的类比，LukF前茎占据与相邻原聚体的氨基闩锁在寡聚毒素中占据的位点大致相同的位点。少

项目成果

N.Sugawara,T.Tomita,T.Sato,and Y.Kamio: "Assembly of Staphylococcus aureus leukocidin into a pore-forming ring-shaped oligomer on human polymorphonuclear leukocytes and rabbit erythrocytes"Biosci.Biotechnol.Biochem.. 63(5). 884-891 (1999)

N.Sukawara、T.Tomita、T.Sato 和 Y.Kamio：“将金黄色葡萄球菌杀白细胞素组装成人多形核白细胞和兔红细胞上的成孔环状寡聚物”Biosci.Biotechnol.Biochem.. 63(5

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

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 φPV83-pro, φPVL, φ11, and other phages"Biosci.Biotechnol.Bi

D. Zou、J. Kaneko、S. Narita 和 Y. Kamio：“金黄色葡萄球菌 P83 染色体上携带 Panton-Valentine 杀白细胞素基因的原噬菌体 φPV83-pro：φPV83-pro、φPVL、 φ11，和其他噬菌体“Biosci.Biotechnol.Bi

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