Presence of a Glycan at a Potential N-Glycosylation Site, Asn-281, of Bovine Lactoferrin

Wei, Zheng; Nishimura, Toshihide; Yoshida, Shigeru

doi:10.3168/jds.s0022-0302(00)74929-0

Cited by 39 publications

(39 citation statements)

References 17 publications

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“…It is N-glycosylated 30 with five potential sites based on the consensus sequence at 252 Asn, 300 Asn, 387 Asn, 495 Asn and 564 Asn. Figure 1a shows the MALDI FT-ICR MS spectrum of b-LF glycopeptides after digestion with pronase in the negative ion mode (Table S-1, supplementary information).…”

Section: Resultsmentioning

confidence: 99%

Enhanced Detection and Identification of Glycopeptides in Negative Ion Mode Mass Spectrometry

Nwosu

Strum

et al. 2010

Anal. Chem.

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A combined MS and MS/MS approach implemented with MALDI FT-ICR MS in the negative ion mode is described for enhanced glycopeptide detection and MS/MS analysis. Positive ion mode mass spectrometry (MS) analysis is widely used for glycopeptide characterization, but the analyses are hampered by potential charge-induced fragmentation of the glycopeptides and poor detection of the glycopeptides harboring sialic acids. Furthermore, tandem MS analysis (MS/MS) via collision-induced dissociation (CID) of glycopeptides in the positive ion mode predominantly yields glycan fragmentation with minimal information to verify the connecting peptide moiety. In this study, glycoproteins such as, bovine lactoferrin (b-LF) for N-glycosylation and kappa casein (k-CN) for O-glycosylation were analyzed in both the positive- and negative ion modes after digestion with bead-immobilized pronase. For the b-LF analysis, 44 potential N-linked glycopeptides were detected in the positive ion mode while 61 potential N-linked glycopeptides were detected in the negative ion mode. By the same token, more O-linked glycopeptides mainly harboring sialic acids from k-CN were detected in the negative ion mode. The enhanced glycopeptide detection allowed improved site-specific analysis of protein glycosylation and superior to positive ion mode detection. Overall, the negative ion mode approach is aimed towards enhanced N- and O-linked glycopeptide detection and to serve as a complementary tool to the positive ion mode MS/MS analysis.

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Section: Resultsmentioning

confidence: 99%

Enhanced Detection and Identification of Glycopeptides in Negative Ion Mode Mass Spectrometry

Nwosu

Strum

et al. 2010

Anal. Chem.

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“…On the other hand, Wei et al found that bovine lactoferrin isoform-is glycosylated by a complex type N-glycan at the fifth potential N-glycosylation site, Asn281. 34) It has been shown that bovine lactoferrin N-glycans contain certain epitopes, including -Gal (Gal(1-3)Gal(1-4)GlcNAc), LacdiNAc (GalNAc(1-4)GlcNAc), and N-glycolylneuraminic acid (Neu5Gc), while these are not found in human lactoferrin N-glycans.…”

Section: Recent Advances In Studies On the Carbohydrate Moieties Omentioning

confidence: 99%

Recent Advances in Studies on Milk Oligosaccharides of Cows and Other Domestic Farm Animals

Urashima

Taufik

Fukuda

et al. 2013

Bioscience, Biotechnology, and Biochemistry

189

142

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“…Furthermore, LF has been shown to have inhibitory effects on tumourigenesis [12,13] and anti-metastatic activity [14][15][16]. LFs contain between 1 and 5 potential glycosylation sites, depending on the species [17][18][19]. In human milk, LF is a highly abundant protein, with 5.3 mg/ml in colostrum and around 1 mg/ml in later stages of the lactation [20].…”

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confidence: 99%

“…Human LF consists of 691 amino acids and contains 3 potential glycosylation sites at Asn138, Asn479, and Asn624; Asn138 and Asn479 are occupied with complex-type N-glycans, whereas Asn624 is usually unoccupied [25,26]. Bovine LF consists of 689 amino acids and has 5 potential glycosylation sites at Asn233, Asn288, Asn368, Asn476, and Asn545; Asn233, Asn368, Asn467, and Asn545 are always occupied, and Asn288 is occupied for about 30% [18,19,27].…”

mentioning

confidence: 99%

Use of Wisteria floribunda agglutinin affinity chromatography in the structural analysis of the bovine lactoferrin N-linked glycosylation

Leeuwen

Schoemaker

Timmer

et al. 2012

Biochimica et Biophysica Acta (BBA) - General Subjects

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Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Background: Over the years, the N-glycosylation of both human and bovine lactoferrin (LF) has been studied extensively, however not all aspects have been studied in as much detail. Typically, the bovine LF complex-type N-glycans include certain epitopes, not found in human LF N-glycans, i.e. Gal(α1-3)Gal(β1-4)GlcNAc (αGal), GalNAc(β1-4) GlcNAc (LacdiNAc), and N-glycolylneuraminic acid (Neu5Gc). The combined presence of complex-type N-glycans, with αGal, LacdiNAc, LacNAc [Gal(β1-4)GlcNAc], Neu5Ac (N-acetylneuraminic acid), and Neu5Gc epitopes, and oligomannose-type N-glycans complicates the high-throughput analysis of such N-glycoprofiles highly. Methods: For the structural analysis of enzymatically released N-glycan pools, containing both LacNAc and LacdiNAc epitopes, a prefractionation protocol based on Wisteria floribunda agglutinin affinity chromatography was developed. The sub pools were analysed by MALDI-TOF-MS and HPLC-FD profiling, including sequential exoglycosidase treatments.Results: This protocol separates the N-glycan pool into three sub pools, with (1) free of LacdiNAc epitopes, (2) containing LacdiNAc epitopes, partially shielded by sialic acid, and (3) containing LacdiNAc epitopes, without shielding by sialic acid. Structural analysis by MALDI-TOF-MS and HPLC-FD showed a complex pattern of oligomannose-, hybrid-, and complex-type di-antennary structures, both with, and without LacdiNAc, αGal and sialic acid. Conclusions: Applying the approach to bovine LF has led to a more detailed N-glycome pattern, including LacdiNAc, αGal, and Neu5Gc epitopes, than was shown in previous studies. General significance: Bovine milk proteins contain glycosylation patterns that are absent in human milk proteins; particularly, the LacdiNAc epitope is abundant. Analysis of bovine milk serum proteins is therefore excessively complicated. The presented sub fractionation protocol allows a thorough analysis of the full scope of bovine milk protein glycosylation. This article is part of a Special Issue entitled Glycoproteomics.

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Presence of a Glycan at a Potential N-Glycosylation Site, Asn-281, of Bovine Lactoferrin

Cited by 39 publications

References 17 publications

Enhanced Detection and Identification of Glycopeptides in Negative Ion Mode Mass Spectrometry

Enhanced Detection and Identification of Glycopeptides in Negative Ion Mode Mass Spectrometry

Recent Advances in Studies on Milk Oligosaccharides of Cows and Other Domestic Farm Animals

Use of Wisteria floribunda agglutinin affinity chromatography in the structural analysis of the bovine lactoferrin N-linked glycosylation

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