193 nm Ultraviolet Photodissociation for the Characterization of Singly Charged Proteoforms Generated by MALDI.
Kevin J ZemaitisMowei ZhouWilliam KewLjiljana Paša-TolićPublished in: Journal of the American Society for Mass Spectrometry (2023)
MALDI imaging allows for the near-cellular profiling of proteoforms directly from microbial, plant, and mammalian samples. Despite detecting hundreds of proteoforms, identification of unknowns with only intact mass information remains a distinct challenge, even with high mass resolving power and mass accuracy. To this end, many supplementary methods have been used to create experimental databases for accurate mass matching, including bulk or spatially resolved bottom-up and/or top-down proteomics. Herein, we describe the application of 193 nm ultraviolet photodissociation (UVPD) for fragmentation of quadrupole isolated singly charged ubiquitin ( m / z 8565) by MALDI-UVPD on a UHMR HF Orbitrap. This platform permitted the high-resolution accurate mass measurement of not just terminal fragments but also large internal fragments. The outlined workflow demonstrates the feasibility of top-down analyses of isolated MALDI protein ions and the potential toward more comprehensive characterization of proteoforms in MALDI imaging applications.
Keyphrases
- mass spectrometry
- high resolution
- liquid chromatography
- gas chromatography
- high performance liquid chromatography
- tandem mass spectrometry
- photodynamic therapy
- high resolution mass spectrometry
- small molecule
- high speed
- microbial community
- ultra high performance liquid chromatography
- deep learning
- big data
- fluorescence imaging
- ms ms
- artificial intelligence