Proteins are critical structural and functional components of biological organisms. A single gene may produce many different molecular forms of proteins via post-transcriptional, post-translational modifications or noncovalent complex formation with other molecules. New advances in mass-spectrometry-based proteomics have enabled direct analysis of intact proteins and native protein complexes. While the more conventional proteomics approaches study proteins “bottom-up” by measuring small peptide fragments, these techniques offer a “top-down” view of proteins at the intact level, allowing us to better understand their structures and functions.
- Supporting the Structural Biology Integrated Research Platform, these resources can measure stoichiometry of protein-protein, protein-nucleic acid, protein-ligand, and protein-metal complexes. The information will help us better understand protein structure and function, including heterogenous and dynamic protein assemblies.
- Supporting the Biomolecular Pathways Integrated Research Platform, these resources can precisely define “proteoforms” at the intact protein level—combinations of post-translational modifications and in vivo proteolytic cleavages.
- Thermo Orbitrap UHMR
- Waters Synapt G2s-I Ion Mobility Time-of-flight
- Thermo Orbitrap Eclipse Tribrid
- Thermo Orbitrap Fusion Lumos
- IsoForma tool for automated quantification of isomeric proteoforms
- PSpecteR application tool and R package for annotation of fragmentation patterns
Tips for success
- Current technology is best suited for targeted analysis on enriched, purified, or recombinant protein samples. New technologies and instrumentation are also being actively developed for more complex samples; please contact your EMSL science point of contact for details.