Do Rodent and Human Brains Have Different N-Glycosylation Patterns?

Albach, Claus; Klein, Roger A.; Schmitz, Brigitte

doi:10.1515/bc.2001.026

Cited by 33 publications

(26 citation statements)

References 39 publications

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“…5) support the notion that the decreases in M r of the MOPR by the A118G or A112G SNP are due to varied N-glycosylation status, which may imply the elimination of one of the N-linked glycans by the SNP. It has been demonstrated that, in general, glycoproteins from the human brain show similar profiles of brain region-specific N-glycans as those from mouse and rat brains [34]. Therefore, what we observed on the MOPRs in the mouse brains is likely to reflect those in the human brain.…”

Section: Discussionsupporting

confidence: 58%

A common single nucleotide polymorphism A118G of the μ opioid receptor alters its N-glycosylation and protein stability

Huang

Chen

Mague

et al. 2011

Biochemical Journal

107

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Synopsis The A118G single nucleotide polymorphism (SNP) of the human mu opioid receptor (hMOPR) gene OPRM1 results in an amino acid substitution (N40D). Subjects homozygous for G118 allele were reported to require higher morphine doses to achieve adequate analgesia and G118 allele was more prevalent among drug abusers. However, changes in the MOPR protein associated with this SNP are unknown. Using a knock-in mouse model (G/G mice) that possesses the equivalent nucleotide/amino acid substitution (N38D; A112G) of the A118G in the hMOPR gene , we investigated N-linked glycosylation status of thalamic and striatal MOPR in G/G mice vs A/A (wildtype) mice. The relative molecular mass (Mr) of MOPR determined with immunoblotting was lower in G/G mice than in A/A mice. Following treatment with PNGase F, which removes all N-linked glycans, both MOPR variants had identical Mr, indicating that this discrepancy was due to a lower level of N-glycosylation of the MOPR in G/G mice. In CHO cells stably expressing hMOPRs, G118/D40-hMOPR had lower Mr than A118/N40-hMOPR, which was similarly due to differential N-glycosylation. Pulse-chase studies revealed that the half-life of the mature form of G118/D40-hMOPR (∼12h) was shorter than that of A118/N40-hMOPR (∼28h). Thus, A118G SNP reduces MOPR N-glycosylation and protein stability.

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Section: Discussionsupporting

confidence: 58%

A common single nucleotide polymorphism A118G of the μ opioid receptor alters its N-glycosylation and protein stability

Huang

Chen

Mague

et al. 2011

Biochemical Journal

107

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“…N-glycans from GPI-anchored CD59 expressed in Chinese hamster ovary (CHO) cells have been shown to consist of bi-, tri-, and tetraantennary complex structures with up to four N-acetyllactosamine extensions while corresponding glycans from the soluble form of the glycoprotein consisted almost entirely of biantennary compounds . Another study on ''brain-type'' glycosylation (absence of the 3-antenna) has been reported (Albach, Klein, & Schmitz, 2001) and glucuronic acid, an unusual constituent of N-glycans has been found in bovine peripheral myelin glycoprotein, P0 ) and Complex Bowes melanoma tissue (Zamze et al, 2001).…”

Section: N-octylglucosidementioning

confidence: 99%

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update covering the period 2001–2002

Harvey¹

2008

Mass Spectrometry Reviews

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This review is the second update of the original review on the application of MALDI mass spectrometry to the analysis of carbohydrates and glycoconjugates that was published in 1999. It covers fundamental aspects of the technique as applied to carbohydrates, fragmentation of carbohydrates, studies of specific carbohydrate types such as those from plant cell walls and those attached to proteins and lipids, studies of glycosyl-transferases and glycosidases, and studies where MALDI has been used to monitor products of chemical synthesis. Use of the technique shows a steady annual increase at the expense of older techniques such as FAB. There is an increasing emphasis on its use for examination of biological systems rather than on studies of fundamental aspects and method development and this is reflected by much of the work on applications appearing in tabular form.

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“…We could show that all six potential N-glycosylation sites of NCAM from adult mouse brain are modified, at least one of these (N-1) only partially. The differential approach employed here should be particularly valuable in the case of highly heterogeneous glycosylations as found for glycoproteins from different cellular systems and species [15,16,17,26,27,31,32,33,34,35,36,37,38,39,40]. Direct MALDI-TOF-MS studies of glycopeptides have shown pronounced heterogeneous structures on single N-glycosylation sites of NCAM [26].…”

Section: Discussionmentioning

confidence: 99%

“…N-1 was not detected in bovine NCAM and neither N-1 nor N-4 could be identified in murine NCAM in the glycosylated or the non-glycosylated form which may be explained by the strategy in these studies employing enrichment of glycopeptides with antibodies against PSA or HNK-1. Most likely N-1 and N-4 carry acidic and neutral glycans of the types as identified in several studies to be conserved between rodent, bovine, and human brain glycoproteins [15,16,17,27,35,36,37,38,39,40]. Such heterogeneities, and additional problems observed by proteolytic and hydrolytic side-reactions upon deglycosylation may complicate the mass spectrometric analysis of glycosylation sites, thus rendering high-resolution MS approaches particularly valuable as a general method to define glycosylation site usage in glycoproteins.…”

Section: Discussionmentioning

confidence: 99%

Identification of N -glycosylation sites of the murine neural cell adhesion molecule NCAM by MALDI-TOF and MALDI-FTICR mass spectrometry

Albach

Damoc

Denzinger

et al. 2004

Analytical and Bioanalytical Chemistry

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Mass spectrometry has been shown in recent years to be a powerful tool to determine accurate molecular masses and sequences of peptides and proteins and post-translational modifications such as glycosylation, phosphorylation, and sulfation. For glycosylation, it has been increasingly recognized to be of pivotal importance to identify whether potential glycosylation sites are actually modified by glycans, because functions of proteins may be modulated or depend on the presence of glycans at specific sites. Several recent reports have established that mass spectrometric techniques such as matrix-assisted laser desorption/ionization or electrospray ionization mass spectrometry (MALDI-TOF or ESI-MS, respectively) with or without preceding HPLC and in combination with PNGase F treatment are suited to analyze whether consensus sequences for N-glycosylation are glycosylated or not. Here we report the mass spectrometric analysis of the six potential N-glycosylation sites of the neural cell adhesion molecule NCAM from adult mouse brain. Unmodified peptides and glycopeptides each carrying a single glycosylation site were generated from NCAM by AspN and trypsin treatment and submitted to reversed-phase HPLC with or without prior enzymatic release of N-glycans. The resulting peptides were analyzed by MALDI-TOF-MS. In addition, high-resolution Fourier transform-ion cyclotron resonance (MALDI-FTICR) mass spectrometry was performed after in-gel deglycosylation and subsequent trypsin digestion. By using these procedures all six consensus sequences were shown to be glycosylated; the observation of an unmodified peptide with the consensus sequence N-1 indicates only partial glycosylation at this site.

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Do Rodent and Human Brains Have Different N-Glycosylation Patterns?

Cited by 33 publications

References 39 publications

A common single nucleotide polymorphism A118G of the μ opioid receptor alters its N-glycosylation and protein stability

A common single nucleotide polymorphism A118G of the μ opioid receptor alters its N-glycosylation and protein stability

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update covering the period 2001–2002

Identification of N -glycosylation sites of the murine neural cell adhesion molecule NCAM by MALDI-TOF and MALDI-FTICR mass spectrometry

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