Preprint Article Version 1 This version is not peer-reviewed

Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains

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. Preprints 2020, 2020020088 (doi: 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 (doi: 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.

Subject Areas

structural mass spectrometry; protein structure; chemical cross-linking; disuccinimidyl glutarate; bis(sulfosuccinimidyl) glutarate; bovine serum albumin

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