Version 1
: Received: 6 February 2020 / Approved: 7 February 2020 / Online: 7 February 2020 (02:54:08 CET)
How to cite:
Fiala, J.; Kukačka, Z.; Novák, P. Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains. Preprints2020, 2020020088. https://doi.org/10.20944/preprints202002.0088.v1
Fiala, J.; Kukačka, Z.; Novák, P. Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains. Preprints 2020, 2020020088. https://doi.org/10.20944/preprints202002.0088.v1
Fiala, J.; Kukačka, Z.; Novák, P. Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains. Preprints2020, 2020020088. https://doi.org/10.20944/preprints202002.0088.v1
APA Style
Fiala, J., Kukačka, Z., & Novák, P. (2020). Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains. Preprints. https://doi.org/10.20944/preprints202002.0088.v1
Chicago/Turabian Style
Fiala, J., Zdeněk Kukačka and Petr Novák. 2020 "Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains" Preprints. https://doi.org/10.20944/preprints202002.0088.v1
Abstract
The combination of chemical cross-linking and mass spectrometry is currently a progressive technology for deriving structural information of proteins and protein complexes. In addition, chemical cross-linking is a powerful tool for stabilizing macromolecular complexes for single particle cryo-electron microscopy. Broad pallets of cross-linking chemistry, currently available for the majority of cross-linking experiments, still rely on the amine-reactive N-hydroxysuccinimide esters targeting mainly N-termini and lysine side chains. These cross-linkers are divided into two groups: water soluble and water insoluble; and research teams prefer one or another speculating on the benefits of their choice. However, the effect of cross-linker polarity on the outcome of cross-linking reaction has never been studied. Herein, we use both polar (bis(sulfosuccinimidyl) glutarate) and non-polar (disuccinimidyl glutarate) cross-linkers and systematically investigated the impact of cross-linker hydrophobicity on resulting distance constraints, using bovine serum albumin as a model protein.
Keywords
structural mass spectrometry; protein structure; chemical cross-linking; disuccinimidyl glutarate; bis(sulfosuccinimidyl) glutarate; bovine serum albumin
Subject
Chemistry and Materials Science, Analytical Chemistry
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
