[논문]Protein folding from heterogeneous unfolded state revealed by time-resolved X-ray solution scattering

Kim, Tae Wu; Lee, Sang Jin; Jo, Junbeom; Kim, Jong Goo; Ki, Hosung; Kim, Chang Woo; Cho, Kwang Hyun; Choi, Jungkweon; Lee, Jae Hyuk; Wulff, Michael; Rhee, Young Min; Ihee, Hyotcherl

doi:10.1073/pnas.1913442117

[해외논문] Protein folding from heterogeneous unfolded state revealed by time-resolved X-ray solution scattering 원문보기

Proceedings of the National Academy of Sciences of the United States of America, v.117 no.26, 2020년, pp.14996 - 15005

Kim, Tae Wu (Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Lee, Sang Jin , Jo, Junbeom (Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Kim, Jong Goo , Ki, Hosung (Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Kim, Chang Woo , Cho, Kwang Hyun (Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Choi, Jungkweon , Lee, Jae Hyuk (Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Wulff, Michael , Rhee, Young Min (Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Ihee, Hyotcherl

Abstract ▼ AI-Helper

SignificanceProtein undergoes a folding pathway to find a three-dimensional structure that expresses its biological function in a living cell. From vast theoretical studies, the funnel-like free-energy landscape has been recognized as a prominent scheme to describe protein folding, but it has been unclear how the dynamic behavior of protein folding is linked to conformational heterogeneity. Here we employ the time-resolved X-ray scattering method combined with systematic structural analysis to track the folding of cytochrome c. The kinetics and structural changes obtained from the data reveal that various protein conformations in the unfolded state take the complex folding pathways, resulting in the stretched exponential kinetics. This study may provide structural insights into fundamental principles governing protein folding.One of the most challenging tasks in biological science is to understand how a protein folds. In theoretical studies, the hypothesis adopting a funnel-like free-energy landscape has been recognized as a prominent scheme for explaining protein folding in views of both internal energy and conformational heterogeneity of a protein. Despite numerous experimental efforts, however, comprehensively studying protein folding with respect to its global conformational changes in conjunction with the heterogeneity has been elusive. Here we investigate the redox-coupled folding dynamics of equine heart cytochrome c (cyt-c) induced by external electron injection by using time-resolved X-ray solution scattering. A systematic kinetic analysis unveils a kinetic model for its folding with a stretched exponential behavior during the transition toward the folded state. With the aid of the ensemble optimization method combined with molecular dynamics simulations, we found that during the folding the heterogeneously populated ensemble of the unfolded state is converted to a narrowly populated ensemble of folded conformations. These observations obtained from the kinetic and the structural analyses of X-ray scattering data reveal that the folding dynamics of cyt-c accompanies many parallel pathways associated with the heterogeneously populated ensemble of unfolded conformations, resulting in the stretched exponential kinetics at room temperature. This finding provides direct evidence with a view to microscopic protein conformations that the cyt-c folding initiates from a highly heterogeneous unfolded state, passes through still diverse intermediate structures, and reaches structural homogeneity by arriving at the folded state.

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