STRUCTURAL ANALYSIS OF PROTEIN FOLDING REACTION UNDER KINETIC CONTROL

直到最近，人们还普遍认为蛋白质折叠是 控制的。 研究？溶解蛋白酶在我们 实验室已经直接挑战了这一假设。 ?-溶解蛋白酶 (? LP）用166个残基的N-末端前区（Pro）合成。 在不存在Pro的情况下，变性aLP在体外折叠成 中间熔融球状状态。 这个中间体实际上 比原生状态更稳定， 慢慢地回到了祖国。 随着Pro的加入， 反应加速超过109。 Pro催化折叠 通过直接稳定折叠过渡态， 促进中间体转化为动力学上 被困的亚稳态自然状态 我们正在解决 的X射线晶体结构LP/Pro产品复合体。 知识 Pro如何与之交互的细节？LP可能提供线索， 它们在折叠过渡态的相互作用。 这反过来将 深入了解过渡态稳定的来源， 促进我们对Pro催化蛋白质作用机制的理解 折页.

Until recently, it had been widely assumed that protein folding is thermodynamically controlled. Studies of ?-lytic protease in our laboratory have directly challenged this hypothesis. ?-lytic protease (?LP) is synthesized with a 166-residue N-terminal pro region (Pro). In the absence of Pro, denatured aLP folds in vitro to an intermediate, molten globule-like state. This intermediate is in fact thermodynamically more stable than the native state and converts very slowly to the native state. With the addition of Pro, the folding reaction is accelerated by more than 109. Pro catalyzes the folding reaction by directly stabilizing the folding transition state, facilitating the conversion of the intermediate to the kinetically trapped, metastable native state. We are in the process of solving the X-ray crystal structure of the ?LP/Pro product complex. Knowledge of the details of how Pro interacts with ?LP may provide clues to their interaction in the folding transition state. This in turn will yield insight into the source of transition state stabilization and advance our understanding of the mechanism of Pro-catalyzed protein folding.