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Influence of cross-linker polarity on selectivity towards lysine side chains

ABSTRACT: Even though the amine reactive BS2G and DSG cross-linkers have the same length of spacer and are based on N-hydroxysuccinimidic group, our data showed that each of them formed preferentially different cross-links. We demonstrated that the choice of cross-linker can have a significant impact on the output data for structural characterization of biomolecules. Using equimolar mixtures of DSG with d6-BS2G, and BS2G with d6-DSG, we established that the polar BS2G preferentially bound to polar regions of modified molecule, whereas non-polar DSG bound to hydrophobic regions. This phenomenon established that the mixture of polar and non-polar cross-linkers acted as an efficient tool for the determination of distance constraints in proteins.

INSTRUMENT(S): solariX

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Blood Serum

SUBMITTER: Jan Fiala

LAB HEAD: Petr Novák

PROVIDER: PXD017299 | Pride | 2020-02-28

REPOSITORIES: Pride

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Influence of cross-linker polarity on selectivity towards lysine side chains.

Fiala Jan J Kukačka Zdeněk Z Novák Petr P

Journal of proteomics 20200220

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 mainl ...[more]

PMID: 32087376

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