Protein identification by 3D OrbiSIMS to facilitate in situ imaging and depth profiling
peer-reviewed · Nature Communications · 2020
peer-reviewed · Nature Communications · 2020. Anna M. Kotowska et al. Label-free protein characterization at surfaces is commonly achieved using digestion and/or matrix…
| Date | 2020-11-17 |
| Type | peer-reviewed |
| Venue | Nature Communications |
| Publisher | Springer Science and Business Media LLC |
| Contribution | downstream-application |
| DOI | 10.1038/s41467-020-19445-x |
| Citations (OpenAlex) | 76 |
| Venue 2-year citedness | 17.60 |
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 Orbitrap TM 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.
Methods and tools
- 3D OrbiSIMS in situ protein identification: Gas cluster ion beam SIMS with Orbitrap analysis fragments surface proteins in situ, and de novo peptide sequencing annotates 16 proteins, enabling protein imaging and depth profiling in skin.