支柱动力269 -残留蛋白酶Savinase决定从15 n-nmr放松测量。
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Remerowski ML, Pepermans哈,Hilbers CW, Van De Ven陆地
支柱动力269 -残留蛋白酶Savinase决定从15 n-nmr放松测量。
欧元。1996 2月1;235 (3):629 - 40。
- PubMed ID
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8654411 (在PubMed]
- 文摘
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骨干Savinase动力学,269残留的枯草杆菌蛋白酶分泌杆菌lentus,研究了利用15 n放松测量来自proton-detected维1 h-15n-nmr光谱学。15 n spin-lattice速率常数(R1),自旋自旋弛豫速率常数(R2),和1 h-15n核奥佛好塞效应(一)测定84%的骨干酰胺15 n核。模范自由形式主义[利帕里,g . &萨博,a (1982) j。化学。Soc。104年,4546 - 4559年)被用来推导广义有序参数值,S2,可以看作衡量运动的振幅picosecond-nanosecond时间表,每个氮氢键向量。附加的条款用于符合数据包括一个有效的内部运动相关时间(taue)和一个交换术语(雷克斯)交换的贡献占R2。整体旋转相关时间(taum)是9.59 + / - 0.02 ns;平均订单参数(S2)是0.90 + / - 0.07,表明刚性结构与Savinase一致的高度的二级结构和三级折叠紧凑。残留S125-S128位于substrate-binding地区代表蛋白质的最长picosecond-nanosecond展品障碍的时间表。这些残留物也表现出显著的交换条件,可能象征microsecond-millisecond时间表上的运动,这也可以影响邻近的芳基取代的苯基环boronic酸抑制剂用于这项研究。 S103 and G219 in the substrate-binding region, represent the longest stretch of protein which exhibits disorder on the picosecond-nanosecond timescale. These residues also exhibit significant exchange terms, possibly indicative of motion on the microsecond-millisecond timescale, which could also be influenced by the proximity of the phenyl ring of the substituted aryl boronic acid inhibitor used in this study. S103 and G219 in the substrate-binding region also show flexibility on the picosecond-nanosecond timescale. There is also significant motion in the turn, G258-T260, of a small solvent-exposed loop region which may make the protein vulnerable autolysis at that point. Some residues in both calcium-binding sites and nearby also show mobility.