Selective fragmentation of the N-glycan moiety and protein backbone of ribonuclease B on an Orbitrap Fusion Lumos Tribrid mass spectrometer

Rapid Communications in Mass Spectrometry : RCM
Shasha LiZhixin Tian

Abstract

The functional study and application of an intact glycoprotein require the structural characterization of both the protein backbone and the glycan moiety; the former has been successfully demonstrated with selective fragmentation of the protein backbone in CID and ExD; whether the latter can be achieved with selective fragmentation of the glycan moiety remains to be explored. RNase B solution was electrosprayed and its intact glycoforms of GlcNAc2 Mann (n = 5-9) with the highest abundance (charge state z = 16) were isolated individually and fragmented using CID, ETD, HCD, ETciD, and EThcD on the Orbitrap Fusion Lumos Tribrid mass spectrometer; the dissociation parameters were optimized for selective fragmentation of the N-glycan moiety and protein backbone as well as high sequence coverage. The obtained spectra were interpreted using the protein and N-glycan database search engines ProteinGoggle and GlySeeker, respectively. With exploration of different dissociation parameters for all the five methods, selective fragmentation of the N-glycan moiety (the protein backbone staying intact) was observed in both HCD and EThcD at low collisional energies, but only a few matched product ions were observed; more comprehensive fragmentat...Continue Reading

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Citations

Oct 29, 2019·Rapid Communications in Mass Spectrometry : RCM·Dapeng ZhouZhixin Tian
May 19, 2020·Journal of Mass Spectrometry : JMS·Qiong WuYukui Zhang
Jul 3, 2020·Molecular & Cellular Proteomics : MCP·Weiqian CaoPengyuan Yang
Feb 9, 2021·Molecular & Cellular Proteomics : MCP·Weiqian CaoPengyuan Yang
Mar 26, 2021·Mass Spectrometry Reviews·Deborah Chang, Joseph Zaia
Jun 25, 2021·Analytical Chemistry·Zhan GaoYuanjiang Pan
Aug 14, 2019·Journal of the American Chemical Society·Christopher J GraySabine L Flitsch
Sep 11, 2020·Journal of the American Society for Mass Spectrometry·Qi WangHao Chen

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