STRUCTURE/DYNAMICS OF CELL WALL PROTEINS BY REDOR NMR

我们提出了新的固体核磁共振实验来确定结构和 固定化非晶态蛋白质的动力学研究 在细胞壁表面。通用的REDOR和TEDOR分析方法 在过去的五年里，我们为准确的 确定核间距离(并因此确定几何和结构) 冷冻冷冻干燥蛋白复合体，并提出了新的 TEDOR-REDOR和TEDOR-TEDOR联合实验是异核的 直接适用于细胞壁蛋白质问题的技术。这个 固体核磁共振对这类工作的灵敏度已经被 建立在大蛋白质的实际应用上。我们现在照例 在50-KD的1微摩尔上进行远程测量 标记的蛋白质复合体。我们提出了具体的新的REDOR/TEDOR固态 实验表征：(L)麦曲宁多肽抗生素在体内 多层脂类分散体；(2)完整细胞中的肽聚糖桥 金黄色葡萄球菌的壁；(3)细胞-细胞内的蛋白质-寡糖复合体 (4)磷脂酶A2复合体的界面位置。 水合双层；和(5)D-乳酸的脂质结合区 脱氢酶。蛋白质-低聚糖的结构结果 除了实验之外，还将通过使用 非接触式原子力显微镜。所有这些实际应用 可能导致新的科学和新的生物技术，后者包括 固体核磁共振在药物生物配伍设计中的应用 还有抗生素。

We propose new solid-state NMR experiments to determine the structure and dynamics of noncrystallizeable proteins which have been immobilized in or on cell-wall surfaces. The versatile REDOR and TEDOR analytical methods that we have developed in the last five years for the accurate determination of internuclear distances (and hence geometry and structure) of freeze-quenched lyophilized protein complexes, and proposed new combined TEDOR-REDOR and TEDOR-TEDOR experiments, are heteronuclear techniques that are directly applicable to cell-wall protein problems. The sensitivity of solid-state NMR for this kind of work already has been established in practical applications on big proteins. We now routinely perform long-range distance measurements on one micromole of a 50-kD labeled protein complex. We propose specific new REDOR/TEDOR solid-state experiments to characterize: (l) magainin peptide antibiotics in multilamellar lipid dispersions; (2) peptidoglycan bridges in intact cell walls of S. aureus; (3) protein-oligosaccharide complexes at cell-cell interfaces; (4) the interfacial site of phospholipase A2 complexes in a hydrated bilayer; and (5) the lipid-binding region of D-lactate dehydrogenase. Structural results from the protein-oligosaccharide experiments will be complemented by direct imaging of the complexes using non-contact atomic force microscopy. All of these practical applications could lead both to new science and new biotechnology, the latter including the use of solid-state NMR to assist in the biorational design of drugs and antibiotics.