How internal cavities destabilize a protein

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Standard

How internal cavities destabilize a protein. / Xue, Mengjun; Wakamoto, Takuro; Kejlberg, Camilla; Yoshimura, Yuichi; Nielsen, Tania Aaquist; Risør, Michael Wulff; Sanggaard, Kristian Wejse; Kitahara, Ryo; Mulder, Frans A.A.

I: Proceedings of the National Academy of Sciences of the United States of America, Bind 116, Nr. 42, 2019, s. 21031-21036.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Xue, M, Wakamoto, T, Kejlberg, C, Yoshimura, Y, Nielsen, TA, Risør, MW, Sanggaard, KW, Kitahara, R & Mulder, FAA 2019, 'How internal cavities destabilize a protein', Proceedings of the National Academy of Sciences of the United States of America, bind 116, nr. 42, s. 21031-21036. https://doi.org/10.1073/pnas.1911181116

APA

Xue, M., Wakamoto, T., Kejlberg, C., Yoshimura, Y., Nielsen, T. A., Risør, M. W., Sanggaard, K. W., Kitahara, R., & 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), 21031-21036. https://doi.org/10.1073/pnas.1911181116

CBE

Xue M, Wakamoto T, Kejlberg C, Yoshimura Y, Nielsen TA, Risør MW, Sanggaard KW, Kitahara R, Mulder FAA. 2019. How internal cavities destabilize a protein. Proceedings of the National Academy of Sciences of the United States of America. 116(42):21031-21036. https://doi.org/10.1073/pnas.1911181116

MLA

Xue, Mengjun o.a.. "How internal cavities destabilize a protein". Proceedings of the National Academy of Sciences of the United States of America. 2019, 116(42). 21031-21036. https://doi.org/10.1073/pnas.1911181116

Vancouver

Xue M, Wakamoto T, Kejlberg C, Yoshimura Y, Nielsen TA, Risør MW o.a. How internal cavities destabilize a protein. Proceedings of the National Academy of Sciences of the United States of America. 2019;116(42):21031-21036. https://doi.org/10.1073/pnas.1911181116

Author

Xue, Mengjun ; Wakamoto, Takuro ; Kejlberg, Camilla ; Yoshimura, Yuichi ; Nielsen, Tania Aaquist ; Risør, Michael Wulff ; Sanggaard, Kristian Wejse ; Kitahara, Ryo ; Mulder, Frans A.A. / How internal cavities destabilize a protein. I: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Bind 116, Nr. 42. s. 21031-21036.

Bibtex

@article{3e780aa379ba40e09741d577be83975a,
title = "How internal cavities destabilize a protein",
abstract = "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⋅{\AA}-3.",
keywords = "high-pressure NMR, protein folding and cooperativity, protein stability, unfolded state",
author = "Mengjun Xue and Takuro Wakamoto and Camilla Kejlberg and Yuichi Yoshimura and Nielsen, {Tania Aaquist} and Ris{\o}r, {Michael Wulff} and Sanggaard, {Kristian Wejse} and Ryo Kitahara and Mulder, {Frans A.A.}",
year = "2019",
doi = "10.1073/pnas.1911181116",
language = "English",
volume = "116",
pages = "21031--21036",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "The National Academy of Sciences of the United States of America",
number = "42",

}

RIS

TY - JOUR

T1 - How internal cavities destabilize a protein

AU - Xue, Mengjun

AU - Wakamoto, Takuro

AU - Kejlberg, Camilla

AU - Yoshimura, Yuichi

AU - Nielsen, Tania Aaquist

AU - Risør, Michael Wulff

AU - Sanggaard, Kristian Wejse

AU - Kitahara, Ryo

AU - Mulder, Frans A.A.

PY - 2019

Y1 - 2019

N2 - 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.

AB - 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.

KW - high-pressure NMR

KW - protein folding and cooperativity

KW - protein stability

KW - unfolded state

UR - http://www.scopus.com/inward/record.url?scp=85073311113&partnerID=8YFLogxK

U2 - 10.1073/pnas.1911181116

DO - 10.1073/pnas.1911181116

M3 - Journal article

C2 - 31570587

AN - SCOPUS:85073311113

VL - 116

SP - 21031

EP - 21036

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 42

ER -