Site-Specific Glycan Microheterogeneity of Inter-Alpha-Trypsin Inhibitor Heavy Chain H4

Chandler, Kevin Brown; Brnakova, Zuzana; Šanda, Miloslav; Wang, Shuo; Stalnaker, Stephanie H.; Bridger, Robert; Zhao, Peng; Wells, Lance; Edwards, Nathan; Goldman, Radoslav

doi:10.1021/pr500394z

Cited by 36 publications

(36 citation statements)

References 60 publications

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“…Intact glycopeptide analysis offers advantages over analysis of released glycans, primarily the ability to obtain protein-and site-specific glycosylation information. For example, there is growing evidence for the occurrence of nonconsensus motif glycosylation (25,(37)(38)(39)(40), as observed on apolipoprotein E in this study. This site was previously identified as formerly glycosylated in a large-scale proteomic study after PNGase F digestion (39).…”

Section: Discussionsupporting

confidence: 64%

Tissue-Specific Glycosylation at the Glycopeptide Level

Medzihradszky

Kaasik

Chalkley

2015

Molecular & Cellular Proteomics

108

107

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This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N-and O-glycosylation data for ϳ130 proteins. Incorporation of different sialic acid variants in both N-and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. The data suggest that protein N-glycosylation differs depending on cellular location. The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. Molecular

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Section: Discussionsupporting

confidence: 64%

Tissue-Specific Glycosylation at the Glycopeptide Level

Medzihradszky

Kaasik

Chalkley

2015

Molecular & Cellular Proteomics

108

107

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“…Furthermore, the occurrence of N-glycans in HEL that are particularly rich in GlcNAc but lack further galactose residues terminating the antennae (in our data we detected glycans ranging up to (Hex) 3 (HexNAc) 9 ) is fully consistent with previously published work reporting on the composition of Nlinked carbohydrates identified in other glycoproteins from hen egg white. 21 The novel GlycoQuest data analysis workflow used in this work proved to be extremely useful in facilitating the automated detection and characterization of low-abundant N-glycosylation in HEL.…”

Section: Journal Of Proteome Researchsupporting

confidence: 93%

Low Abundant N-linked Glycosylation in Hen Egg White Lysozyme Is Localized at Nonconsensus Sites

et al. 2015

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Although wild-type hen egg white lysozyme (HEL) is lacking the consensus sequence motif NX(S/T), in 1995 Trudel et al. (Biochem. Cell Biol. 1995, 73, 307-309) proposed the existence of a low abundant N-glycosylated form of HEL; however, the identity of active glycosylation sites in HEL remained a matter of speculation. For the first time since Trudel's initial work, we report here a comprehensive characterization by means of mass spectrometry of N-glycosylation in wild-type HEL. Our analytical approach comprised ZIC-HILIC enrichment of N-glycopeptides from HEL trypsin digest, deglycosylation by (18)O/PNGase F as well as by various endoglycosidases, and LC-MS/MS analysis of both intact and deglycosylated N-glycopeptides engaging multiple techniques of ionization and fragmentation. A novel data interpretation workflow based on MS/MS spectra classification and glycan database searching enabled the straightforward identification of the asparagine-rich N-glycopeptide [34-45] FESNFNTQATNR and allowed for compositional profiling of its modifying N-glycans. The overall heterogeneity profile of N-glycans in HEL comprised at least 26 different compositions. Results obtained from deglycosylation experiments provided clear evidence of asparagine residues N44 and N39 representing active glycosylation sites in HEL. Both of these sites do not fall into any known N-glycosylation-specific sequence motif but are localized in rarely observed nonconsensus sequons (NXN, NXQ).

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“…It is a positive APP regulated by IL-6 and correlated with liver development and regeneration (Chandler et al, 2014). Alpha-2-macroglobulin (A2 M), which is a protein in the alpha macroglobulin family that is related to the innate immune system, is responsible for mediating proliferation of T cells and macrophages, as well as acting as an inhibitor of nonspecific endogenous and exogenous proteases (Wang et al, 2012).…”

Section: Discussionmentioning

confidence: 99%