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Ultraviolet Photodissociation of ESI and MALDI generated protein ions on a Q-Exactive mass spectrometer

ABSTRACT: UVPD was implemented on an Orbitrap Q-Exactive plus equipped with a ESI/EP-MALDI. UVPD of MALDI generated ions was benchmarked against MALDI-ISD, MALDI-HCD and ESI-UVPD. MALDI UVPD outperformed MALDI HCD and ISD efficiently sequencing small proteins ions.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Equus Caballus (horse) Homo Sapiens (human)

SUBMITTER: Marialaura Dilillo

LAB HEAD: Liam A. McDonnell

PROVIDER: PXD011526 | Pride | 2019-01-24

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	ESI-9.5Torr-TB4-z7_HCD25-qb.pcml	Other
	ESI-9.5Torr-TB4-z7_HCD25-qb.raw	Raw
	ESI-9.5Torr-TB4-z7_HCD30-qb.pcml	Other
	ESI-9.5Torr-TB4-z7_HCD30-qb.raw	Raw
	ESI-9.5Torr-TB4-z7_UVPD1pt5-qb.pcml	Other

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Publications

Ultraviolet Photodissociation of ESI- and MALDI-Generated Protein Ions on a Q-Exactive Mass Spectrometer.

Dilillo Marialaura M de Graaf Erik L EL Yadav Avinash A Belov Mikhail E ME McDonnell Liam A LA

Journal of proteome research 20181204 1

The identification of molecular ions produced by MALDI or ESI strongly relies on their fragmentation to structurally informative fragments. The widely diffused fragmentation techniques for ESI multiply charged ions are either incompatible (ECD and ETD) or show lower efficiency (CID, HCD), with the predominantly singly charged peptide and protein ions formed by MALDI. In-source decay has been successfully adopted to sequence MALDI-generated ions, but it further increases spectral complexity, and ...[more]

PMID: 30484663

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Project description:We analyzed the backbone fragmentation behavior of tryptic peptides of a four protein mixture and of E. coli lysate subjected to Ultraviolet Photodissociation (UVPD) at 213 nm on a commercially available UVPD-equipped tribrid mass spectrometer. We obtained 15,178 high-confidence peptide-spectrum matches by additionally recording a reference beam-type collision-induced dissociation (HCD) spectrum of each precursor. Type a, b and y ions were most prominent in UVPD spectra and median sequence coverage ranged from 5.8% (at 5 ms laser excitation time) to 45.0% (at 100 ms). Overall sequence fragment intensity remained relatively low (median: 0.4% (5 ms) to 16.8% (100 ms) of total intensity) and remaining precursor intensity high. Sequence coverage and sequence fragment intensity ratio correlated with precursor charge density, suggesting that UVPD at 213 nm may suffer from newly formed fragments sticking together due to non-covalent interactions. UVPD fragmentation efficiency therefore might benefit from supplemental activation, as was shown for ETD. Aromatic amino acids, most prominently tryptophan, facilitated UVPD. This points at aromatic tags as possible enhancers of UVPD. Data are available via ProteomeXchange and on spectrumviewer.org/db/UVPD_213nm_trypPep

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Project description:In this study we benchmark the performance of ultraviolet photodissociation (UVPD) using 213 nm photons generated by a solid-state laser applied to the study of intact proteoforms from three organisms. 213 nm UVPD was compared to the commonly employed higher-energy collisional dissociation (HCD).

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