Facile Determination of Phosphorylation Sites in Peptides Using Two-Dimensional Mass Spectrometry.
Johanna ParisTomos E MorganChristopher A WoottonMark P BarrowJohn O'HaraPeter B O'ConnorPublished in: Analytical chemistry (2020)
Detection and characterization of phosphopeptides by infrared multiphoton dissociation two-dimensional mass spectrometry (IRMPD 2DMS) is shown to be particularly effective. A mixture of phosphopeptides was analyzed by 2DMS without any prior separation. 2DMS enables the data independent analysis of the mixture and the correlation of the fragments to their precursor ions. The extraction of neutral loss lines corresponding to the loss of phosphate under IRMPD fragmentation allows the selective identification of phosphopeptides. Resonance of the 10.6 μm infrared radiation with the vibrational modes of the phosphate functional group produced efficient absorption and high cleavage coverage of the phosphopeptides at much lower irradiation fluence than for nonphosphorylated peptides improving discrimination. Additionally, the localization of the phosphate group was determined.
Keyphrases
- mass spectrometry
- liquid chromatography
- energy transfer
- quantum dots
- gas chromatography
- capillary electrophoresis
- high performance liquid chromatography
- solid phase extraction
- high resolution
- tandem mass spectrometry
- amino acid
- electronic health record
- radiation induced
- healthcare
- density functional theory
- big data
- molecular dynamics simulations
- dna binding
- real time pcr
- label free
- molecularly imprinted
- simultaneous determination
- electron transfer
- molecular dynamics
- affordable care act
- reduced graphene oxide
- sensitive detection
- gold nanoparticles