Structures of Staphylococcus aureus cell-wall complexes with vancomycin, eremomycin, and chloroeremomycin derivatives by 13C{19F} and 15N{19F} rotational-echo double resonance
Structures of Staphylococcus aureus cell-wall complexes with vancomycin, eremomycin, and chloroeremomycin derivatives by 13C{19F} and 15N{19F} rotational-echo double resonance
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DOI：
10.1021/bi052660s
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发表时间：
2006-04-25
期刊：
影响因子：
2.9
通讯作者：
Schaefer, J
中科院分区：
文献类型：
--
作者：
Kim, SJ;Cegelski, L;Schaefer, J
固态核磁共振已被用来检查分离的细胞壁和完整的金黄色葡萄球菌的完整细胞复合五种不同的万古霉素，伊瑞霉素，氯伊瑞霉素衍生物。用D-[1-C-13]丙氨酸或[1-C-13]甘氨酸和[ε-N-15]赖氨酸的组合特异性标记细胞壁和全细胞。每个结合的糖肽在其C-末端或其二糖位置具有F-19标记的取代基。连接糖肽复合的肽聚糖茎与其相邻茎的细胞壁C-13标记的桥接五糖基区段的C-13{F-19}旋转回波双共振（REDOR）移相指示所有结合的糖肽的氟标记位于桥的一端或另一端。一个例外是N '-（对三氟甲氧基苄基）氯伊瑞霉素，其疏水取代基的长度与奥利万星N'-4-[（4-氯苯基）苄基]氯伊瑞霉素相比相差一个苯基。对于这种药物，氟标记靠近五甘氨酰片段的中间。N-15{F-19} REDOR移相显示，对于C-末端取代的部分，氟接近于五甘氨酰基桥的桥连接位点，对于双硫醚取代的部分，氟接近于交联位点。用D-[1-C-13]丙氨酸标记的细胞壁复合物的全回波R-EDOR光谱（在丙氨酸消旋酶抑制剂存在下）揭示了三种不同的羰基碳化学位移环境，它们来自D-Ala-D-Ala结合位点和D-Ala-Gly- I交联位点。REDOR结果表明每个肽聚糖复合物中有一个氟移相中心。与从C-13{F-19}和N-15{F-19} REDOR失相获得的核间距一致的成熟细胞壁复合物的分子模型允许糖肽的结构和抗菌活性的一致性。
Solid-state NMR has been used to examine isolated cell walls and intact whole cells of Staphylococcus aureus complexed to five different vancomycin, eremomycin, and chloroeremomycin derivatives. The cell walls and whole cells were specifically labeled with D-[1-C-13]alanine, or a combination of [1-C-13]glycine and [epsilon-N-15]lysine. Each of the bound glycopeptides had a F-19-labeled substituent at either its C-terminus or its disaccharide position. The C-13{F-19} rotational-echo double-resonance (REDOR) dephasing for the cell-wall C-13-labeled bridging pentaglycyl segment connecting a glycopeptide-complexed peptidoglycan stern with its neighboring stem indicates that the fluorine labels for all bound glycopeptides are positioned at one or the other end of the bridge. An exception is N'-(p-trifluoromethoxybenzyl)chloroeremomycin, whose hydrophobic substituent differs in length by one phenyl group compared to that of oritavancin, N'-4-[(4-chlorophenyl)benzyl]chloroeremomycin. For this drug, the fluorine label is near the middle of the pentaglycyl segment. N-15{F-19} REDOR dephasing shows the proximity of the fluorine to the bridge-link site of the pentaglycyl bridge for C-terminus-substituted moieties and the crosslink site for disaccharide-substituted moieties. Full-echo R-EDOR spectra of cell-wall complexes from cells labeled by D-[1-C-13]alanine (in the presence of an alanine racemase inhibitor) reveal three different carbonyl carbon chemical-shift environments, arising from the D-Ala-D-Ala binding site and the D-Ala-Gly- I cross-link site. The REDOR results indicate a single fluorine dephasing center in each peptidoglycan complex. Molecular models of the mature cell-wall complexes that are consistent with internuclear distances obtained from C-13{F-19} and N-15{F-19} REDOR dephasing allow a con-elation of structure and antimicrobial activity of the glycopeptides.