Protein identification by 3D OrbiSIMS to facilitate in situ imaging and depth profiling

Kotowska, Anna M.; Trindade, Gustavo F.; Mendes, Paula M.; Williams, Philip M.; Aylott, Jonathan W.; Shard, Alexander G.; Alexander, Morgan R.; Scurr, David J.

Abstract

Label-free protein characterization at surfaces is commonly achieved using digestion and/or matrix application prior to mass spectrometry. We report the assignment of undigested proteins at surfaces in situ using secondary ion mass spectrometry (SIMS). Ballistic fragmentation of proteins induced by a gas cluster ion beam (GCIB) leads to peptide cleavage producing fragments for subsequent OrbitrapTM analysis. In this work we annotate 16 example proteins (up to 272 kDa) by de novo peptide sequencing and illustrate the advantages of this approach by characterizing a protein monolayer biochip and the depth distribution of proteins in human skin.

Citation

Kotowska, A. M., Trindade, G. F., Mendes, P. M., Williams, P. M., Aylott, J. W., Shard, A. G., Alexander, M. R., & Scurr, D. J. (2020). Protein identification by 3D OrbiSIMS to facilitate in situ imaging and depth profiling. Nature Communications, 11(1), Article 5832. https://doi.org/10.1038/s41467-020-19445-x

• Journal Article Type: Article
• Acceptance Date: Oct 9, 2020
• Online Publication Date: Nov 17, 2020
• Publication Date: 2020-12
• Deposit Date: Oct 16, 2020
• Publicly Available Date: Nov 18, 2020
• Journal: Nature Communications
• Electronic ISSN: 2041-1723
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 1
• Article Number: 5832
• DOI: https://doi.org/10.1038/s41467-020-19445-x
• Keywords: General Biochemistry, Genetics and Molecular Biology; General Physics and Astronomy; General Chemistry
• Public URL: https://nottingham-repository.worktribe.com/output/4967341
• Publisher URL: https://www.nature.com/articles/s41467-020-19445-x