Structural Mass Spectrometry of Protein Assemblies

Cross-linking

In theory:

Cross-linking is now commonly used for structural purposes. Briefly, a cross-linker is used to link reactive amino-acids that are close together. After proteolytic digestion, the cross-linked peptides can be enriched (ion-exchange, SEC, biotinylated cross-linkers) and analysed by LC-MSMS. The distance restraints obtained by localizing the cross-linked residues can be used as an input to refine structural models.

In Practice:

We are currently implementing cross-linking MS at the IPBS on our orbitraps.

References

2018

Trcka F, Durech M, Vankova P, Chmelík J, Martinkova V, Hausner J, Kadek A, Marcoux J, Klumpler T, Vojtesek B, Muller P, Man P. “Human stress-inducible Hsp70 has a high propensity to form ATP-dependent antiparallel dimers that are differentially regulated by co-chaperone binding” Molecular & Cellular Proteomics In Press doi: 10.1074/mcp.RA118.001044.

2015

Chabrol E, Thépaut M, Dezutter-Dambuyant C, Vivès C, Marcoux J, Kahn R, Valadeau J, Vachette P, Durand D, Fieschi F. “Alteration of the Langerin oligomerisation state affects Birbeck Granule formation” Biophysical Journal 108(3) : 666-677.

 2014

Marcoux J, Politis A, Marshall D, Rinehart D, Wallace MI, Tamm LK, Robinson CV. “A new model for full length OmpA: native mass spectrometry reveals partial dimerization via C-ter domain” Structure 22(5) : 781-790.

 2013

Rozbeský D, Sovová Z, Marcoux J, Man P, Ettrich R, Robinson CV, Novák P. “Structural model of lymphocyte receptor NKR-P1C revealed by mass spectrometry and molecular modelling” Analytical Chemistry 85(3) : 1597-604.