Protein identification by 3D OrbiSIMS to facilitate in situ imaging and depth profiling.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 33203841.
- Also identified by DOI 10.1038/s41467-020-19445-x and PMC identifier 7672064.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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<sup>TM</sup> 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.
Medical subject headings
- Proteins
- Proteomics
- Skin
- Spectrometry, Mass, Secondary Ion