Ultrarapid mixing experiments shed new light on the characteristics of the initial conformational ensemble during the folding of ribonuclease A.
Ultrarapid mixing experiments shed new light on the characteristics of the initial conformational ensemble during the folding of ribonuclease A.
复制标题
超快速混合实验为核糖核酸酶 A 折叠过程中初始构象整体的特征提供了新的线索。
DOI：
10.1073/pnas.0407999101
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Roder,Heinrich
中科院分区：
文献类型：
--
作者：
Welker,Ervin;Maki,Kosuke;Shastry,MCRamachandra;Juminaga,Darmawi;Bhat,Rajiv;Scheraga,HaroldA;Roder,Heinrich
The earliest folding events in single-tryptophan mutants of RNase A were investigated by fluorescence measurements by using a combination of stopped-flow and continuous-flow mixing experiments covering the time range from 70 μs to 10 s. An ultrarapid double-jump mixing protocol was used to study refolding from an unfolded ensemble containing only native proline isomers. The continuous-flow measurements revealed a series of kinetic events on the submillisecond time scale that account for the burst-phase signal observed in previous stopped-flow experiments. An initial increase in fluorescence within the 70-μs dead time of the continuous-flow experiment is consistent with a relatively nonspecific collapse of the polypeptide chain whereas a subsequent decrease in fluorescence with a time constant of ≈80 μs is indicative of a more specific structural event. These rapid conformational changes are not observed if RNase A is allowed to equilibrate under denaturing conditions, resulting in formation of nonnative proline isomers. Thus, contrary to previous expectations, the isomerization state of proline peptide bonds can have a major impact on the structural events during early stages of folding.
登录
查看更多内容
影响因子：
3.8
作者：
MASKELL, DJ;SWEENEY, KJ;DOUGAN, G
通讯作者：
DOUGAN, G
影响因子：
3.5
作者：
CLOSE, TJ;RODRIGUEZ, RL
通讯作者：
RODRIGUEZ, RL
转座子 γ δ 在克隆基因分析中的用途。
影响因子：
--
作者：
M. Guyer
通讯作者：
M. Guyer
一系列带有各种耐药标记的 Tn5 变体和用于转座子诱变的自杀载体。
影响因子：
3.5
作者：
Sasakawa Chihiro;Yoshikawa Masanosuke
通讯作者：
Yoshikawa Masanosuke
影响因子：
3.7
作者：
LENNOX, ES
通讯作者：
LENNOX, ES