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@inbook{2374292,
author = {Uhrik, Lukas and Henek, Tomas and Planas Iglesias, Joan and Kucera, Josef and Damborský, Jiří and Marek, Martin and Hernychova, Lenka},
address = {New York},
booktitle = {Advanced Methods in Structural Biology},
doi = {http://dx.doi.org/10.1007/978-1-0716-3147-8_18},
editor = {Ângela Sousa, Luis Passarinha},
keywords = {Ancestral luciferase; Hydrogen/deuterium exchange; LoopGrafter; Mass spectrometry; Protein dynamics; Protein engineering},
howpublished = {tištěná verze "print"},
language = {eng},
location = {New York},
isbn = {978-1-0716-3146-1},
pages = {293-318},
publisher = {Springer Nature},
title = {Study of Protein Conformational Dynamics Using Hydrogen/Deuterium Exchange Mass Spectrometry},
url = {https://link.springer.com/protocol/10.1007/978-1-0716-3147-8_18},
year = {2023}
}
TY - CHAP
ID - 2374292
AU - Uhrik, Lukas - Henek, Tomas - Planas Iglesias, Joan - Kucera, Josef - Damborský, Jiří - Marek, Martin - Hernychova, Lenka
PY - 2023
TI - Study of Protein Conformational Dynamics Using Hydrogen/Deuterium Exchange Mass Spectrometry
VL - 1.
PB - Springer Nature
CY - New York
SN - 9781071631461
KW - Ancestral luciferase
KW - Hydrogen/deuterium exchange
KW - LoopGrafter
KW - Mass spectrometry
KW - Protein dynamics
KW - Protein engineering
UR - https://link.springer.com/protocol/10.1007/978-1-0716-3147-8_18
N2 - Intrinsic protein dynamics contribute to their biological functions. Rational engineering of protein dynamics is extremely challenging with only a handful of successful examples. Hydrogen/deuterium exchange coupled to mass spectrometry (HDX-MS) represents a powerful technique for quantitative analysis of protein dynamics. Here we provide a detailed description of the preparation of protein samples, collection of high-quality data, and their in-depth analysis using various computational tools. We illustrate the application of HDX-MS for the study of protein dynamics in the rational engineering of flexible loops in the reconstructed ancestor of haloalkane dehalogenase and Renilla luciferase. These experiments provided unique and valuable data rigorously describing the modification of protein dynamics upon grafting of the loop-helix element. Tips and tricks are provided to stimulate the wider use of HDX-MS to study and engineer protein dynamics.
ER -
UHRIK, Lukas, Tomas HENEK, Joan PLANAS IGLESIAS, Josef KUCERA, Jiří DAMBORSKÝ, Martin MAREK and Lenka HERNYCHOVA. Study of Protein Conformational Dynamics Using Hydrogen/Deuterium Exchange Mass Spectrometry. In \^Angela Sousa, Luis Passarinha. \textit{Advanced Methods in Structural Biology}. New York: Springer Nature, 2023, p.~293-318. 1. ISBN~978-1-0716-3146-1. Available from: https://dx.doi.org/10.1007/978-1-0716-3147-8\_{}18.
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