FOLDING MECHANISMS OF MULTISUBUNIT PEPTIDES AND PROTEINS

这是一项更新建议(5-10年)，以继续研究 多亚单位蛋白质的折叠和组装机制。具体地说， 需要解决的问题是：(1)重要的结构特征是什么 观察到的瞬时中间体和中间过渡态 在特定的代表性多聚体蛋白的折叠过程中？(2) 单体中预先形成的结构影响二聚体的折叠动力学？ (3)部分折叠的伴生物种是否将复性反应引导到 天然二聚体(三聚体)？生物物理技术的组合跨越了 诱变、氢-氚交换、核磁共振、质谱学和 将使用快速混合光学技术来获取关于 折叠过程中随时间变化的结构形成和缔合反应 三个不同的系统：(A)盘绕线圈，(B)色氨酸抑制物和(C) 三聚体β-螺旋蛋白。

This is a renewal proposal (years 5-10) to continue studies of the folding and assembly mechanisms in multisubunit proteins. Specifically, the questions to be addressed are: (1) What are the important structural features of the transient intermediates and intervening transition states observed during the folding of specific representative multimeric protein? (2) Does pre-formed structure in the monomer affect the folding kinetics of the dimer? (3) Do partially folded associated species guide the refolding reaction to the native dimer (trimer)? A combination of biophysical techniques spanning mutagenesis, hydrogen-deuterium exchange, NMR, Mass Spectrometry and rapid-mixing optical techniques will be used to gain information on the time-dependent structure formation and association reactions during the folding of three distinct systems: (a) coiled-coils, (b) the trp-repressor and (c) the trimeric beta-helix proteins.