Detailed Information on Publication Record
2023
Study of Protein Conformational Dynamics Using Hydrogen/Deuterium Exchange Mass Spectrometry
UHRIK, Lukas, Tomas HENEK, Joan PLANAS IGLESIAS, Josef KUCERA, Jiří DAMBORSKÝ et. al.Basic information
Original name
Study of Protein Conformational Dynamics Using Hydrogen/Deuterium Exchange Mass Spectrometry
Authors
UHRIK, Lukas, Tomas HENEK, Joan PLANAS IGLESIAS (724 Spain, belonging to the institution), Josef KUCERA, Jiří DAMBORSKÝ (203 Czech Republic, belonging to the institution), Martin MAREK (203 Czech Republic, belonging to the institution) and Lenka HERNYCHOVA
Edition
New York, Advanced Methods in Structural Biology, p. 293-318, 26 pp. 1. 2023
Publisher
Springer Nature
Other information
Language
English
Type of outcome
Kapitola resp. kapitoly v odborné knize
Field of Study
30403 Technologies involving identifying the functioning of DNA, proteins and enzymes and how they influence the onset of disease and maintenance of well-being
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
Publication form
printed version "print"
References:
RIV identification code
RIV/00216224:14310/23:00133541
Organization unit
Faculty of Science
ISBN
978-1-0716-3146-1
Keywords in English
Ancestral luciferase; Hydrogen/deuterium exchange; LoopGrafter; Mass spectrometry; Protein dynamics; Protein engineering
Tags
Tags
International impact, Reviewed
Změněno: 15/2/2024 14:48, Mgr. Marie Šípková, DiS.
Abstract
V originále
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.
Links
GA22-09853S, research and development project |
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LX22NPO5102, research and development project |
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