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DOI	10.1073/pnas.1911181116
	How internal cavities destabilize a protein
	Xue M.; Wakamoto T.; Kejlberg C.; Yoshimura Y.; Nielsen T.A.; Risør M.W.; Sanggaard K.W.; Kitahara R.; Mulder F.A.A.
发表日期	2019
ISSN	0027-8424
起始页码	21031
结束页码	21036
卷号	116 期号:42
英文摘要	Although many proteins possess a distinct folded structure lying at a minimum in a funneled free energy landscape, thermal energy causes any protein to continuously access lowly populated excited states. The existence of excited states is an integral part of biological function. Although transitions into the excited states may lead to protein misfolding and aggregation, little structural information is currently available for them. Here, we show how NMR spectroscopy, coupled with pressure perturbation, brings these elusive species to light. As pressure acts to favor states with lower partial molar volume, NMR follows the ensuing change in the equilibrium spectroscopically, with residue-specific resolution. For T4 lysozyme L99A, relaxation dispersion NMR was used to follow the increase in population of a previously identified “invisible” folded state with pressure, as this is driven by the reduction in cavity volume by the flipping-in of a surface aromatic group. Furthermore, multiple partly disordered excited states were detected at equilibrium using pressure-dependent H/D exchange NMR spectroscopy. Here, unfolding reduced partial molar volume by the removal of empty internal cavities and packing imperfections through subglobal and global unfolding. A close correspondence was found for the distinct pressure sensitivities of various parts of the protein and the amount of internal cavity volume that was lost in each unfolding event. The free energies and populations of excited states allowed us to determine the energetic penalty of empty internal protein cavities to be 36 cal·Å−3 © 2019 National Academy of Sciences. All rights reserved.
英文关键词	High-pressure NMR; Protein folding and cooperativity; Protein stability; Unfolded state
语种	英语
scopus关键词	lysozyme; protein; Article; controlled study; nuclear magnetic resonance spectroscopy; priority journal; protein domain; protein folding; protein stability; protein unfolding; chemistry; Enterobacteria phage T4; nuclear magnetic resonance; pressure; procedures; protein conformation; protein denaturation; Bacteriophage T4; Muramidase; Nuclear Magnetic Resonance, Biomolecular; Pressure; Protein Conformation; Protein Denaturation; Protein Folding; Proteins
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160340
作者单位	Xue, M., Interdisciplinary Nanoscience Center, University of Aarhus, Aarhus C, 8000, Denmark, Department of Chemistry, University of Aarhus, Aarhus C, 8000, Denmark, Department of Chemistry, University of Washington, Seattle, WA 98195, United States; Wakamoto, T., Graduate School of Life Sciences, Ritsumeikan University, Shiga, 525-8577, Japan; Kejlberg, C., Interdisciplinary Nanoscience Center, University of Aarhus, Aarhus C, 8000, Denmark, Department of Chemistry, University of Aarhus, Aarhus C, 8000, Denmark; Yoshimura, Y., Interdisciplinary Nanoscience Center, University of Aarhus, Aarhus C, 8000, Denmark, Department of Chemistry, University of Aarhus, Aarhus C, 8000, Denmark, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima City, Hiroshima, 739-8526, Japan; Nielsen, T.A., Interdisciplinary Nanoscience Center, University of Aarhus, Aarhus C, 8000, Denmark, Department of Chemistry, University of Aarhus, Aarhus C, 8000, Denmark; Risør, M.W., Interdisciplinary Nan...
推荐引用方式 GB/T 7714	Xue M.,Wakamoto T.,Kejlberg C.,et al. How internal cavities destabilize a protein[J],2019,116(42).
APA	Xue M..,Wakamoto T..,Kejlberg C..,Yoshimura Y..,Nielsen T.A..,...&Mulder F.A.A..(2019).How internal cavities destabilize a protein.Proceedings of the National Academy of Sciences of the United States of America,116(42).
MLA	Xue M.,et al."How internal cavities destabilize a protein".Proceedings of the National Academy of Sciences of the United States of America 116.42(2019).