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2013 ; 85
(12
): 5666-75
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gab.com Text
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Automated assignments of N- and O-site specific glycosylation with extensive
glycan heterogeneity of glycoprotein mixtures
#MMPMID23662732
Strum JS
; Nwosu CC
; Hua S
; Kronewitter SR
; Seipert RR
; Bachelor RJ
; An HJ
; Lebrilla CB
Anal Chem
2013[Jun]; 85
(12
): 5666-75
PMID23662732
show ga
Site-specific glycosylation (SSG) of glycoproteins remains a considerable
challenge and limits further progress in the areas of proteomics and glycomics.
Effective methods require new approaches in sample preparation, detection, and
data analysis. While the field has advanced in sample preparation and detection,
automated data analysis remains an important goal. A new bioinformatics approach
implemented in software called GP Finder automatically distinguishes correct
assignments from random matches and complements experimental techniques that are
optimal for glycopeptides, including nonspecific proteolysis and high mass
resolution liquid chromatography/tandem mass spectrometry (LC/MS/MS). SSG for
multiple N- and O-glycosylation sites, including extensive glycan heterogeneity,
was annotated for single proteins and protein mixtures with a 5% false-discovery
rate, generating hundreds of nonrandom glycopeptide matches and demonstrating the
proof-of-concept for a self-consistency scoring algorithm shown to be compliant
with the target-decoy approach (TDA). The approach was further applied to a
mixture of N-glycoproteins from unprocessed human milk and O-glycoproteins from
very-low-density-lipoprotein (vLDL) particles.