Mass spectrometry of apolipoprotein C-III, a simple analytical method for mucin-type O-glycosylation and its application to an autosomal recessive cutis laxa type-2 (ARCL2) patient

Wada, Yoshinao; Kadoya, Machiko; Okamoto, Nobuhiko

doi:10.1093/glycob/cws086

Cited by 40 publications

(56 citation statements)

References 39 publications

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“…Sample preparation for MALDI analysis of native apoC‐III was performed as previously described in detail . Briefly, 2 μl of serum samples were diluted in 50 μl of 0.1% trifluoroacetic acid, then subjected to prolonged agitation by vortex (15 min) and desalted by ZipTip C18 (Merck Millipore, Darmstadt).…”

Section: Methodssupporting

confidence: 91%

“…Species were detected as [M+H] + molecular ions (average masses) in their oxidized form (ox). Minor apoC‐III forms lacking alanine terminal residue(s) because of carboxypeptidase activity may be also present as previously described . Glycan structures were assigned following consortium for functional glycomics guidelines: N ‐acetylgalactosamine, yellow square; galactose, yellow circle; sialic acid, purple lozenge.…”

Section: Resultsmentioning

confidence: 98%

“…One of the advantages of profiling apoC‐III O ‐glycosylation by MALDI‐MS is the possibility to distinguish, among the asialo‐glycoforms (apoC‐III 0 ), aglycosylated species (apoC‐III 0a ) from glycosylated species bearing only a GalNAc residue (apoC‐III 0b ) or a disaccharide glycan moiety Gal‐GalNAc (apoC‐III 0c ). This is particularly important when dealing with CDG …”

Section: Introductionmentioning

confidence: 99%

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MALDI‐MS profiling of serum O‐glycosylation and N‐glycosylation in COG5‐CDG

et al. 2017

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Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III (aglycosylated glycoform), whereas apoC-III (mono-sialylated glycoform) decreased significantly. Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. Copyright © 2017 John Wiley & Sons, Ltd.

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Section: Methodssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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MALDI‐MS profiling of serum O‐glycosylation and N‐glycosylation in COG5‐CDG

et al. 2017

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“…26-28 The applicability of these methods to investigate aberrations in glycosylation associated with disease samples has been demonstrated. 29-31 Enrichment techniques, such as lectin affinity chromatography, or hydrazide chemistry, and peptide labeling have also been used in conjunction with LC-MS/MS to characterize and quantify the glycosylation sites of glycoproteins. 32-34 …”

Section: Introductionmentioning

confidence: 99%

LC–MS/MS Identification of the O-Glycosylation and Hydroxylation of Amino Acid Residues of Collagen α-1 (II) chain from Bovine Cartilage

Song

Mechref

2013

J. Proteome Res.

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O-Glycosylation of collagen is a unique type of posttranslational modifications (PTMs), involving the attachment of galactose (Gal) or glucose-galactose (Glc-Gal) moieties to hydroxylysine (HyK). Also, hydroxyproline (HyP) result from the posttranslational hydroxylation of some proline residues in collagen. Here, LC-MS/MS was effectively employed to identify 23 O-glycosylation sites and a large number of HyP residues associated with bovine type II collagen α-1 chain (CO2A1). The modifications of the 23 O-glycosylation sites varied qualitatively and quantitatively. Both Gal and Glc-Gal moieties occupied 22 of the identified glycosylation sites while K773 was observed as unmodified. A large number of HyP residues at Yaa positions of Gly-Xaa-Yaa motif were detected. HyP residues at Xaa positions of Gly-HyP-HyP, Gly-HyP-Ala, and Gly-HyP-Val motifs were also observed. Notably, HyP residue of Gly-HyP-Gln motif was detected, which has not been previously reported. Moreover, the deamidation of 8 Asn residues was identified, of which 2 Asp residues were observed at different retention times because of isomerization (Asp vs. isoAsp). Partial macroheterogeneities of some CO2A1 glycosylation sites were revealed by LC-MS/MS analysis. ETD experiments revealed partial macroheterogeneities associated with K299-K308, K452-K464, K464-K470 and K857-K884 glycosylation sites. Semi-quantitative data suggest that the glycosylation of hydroxylysine residues is site-specific.

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“…ApoC-III is highly heterogeneous protein because of the extensive protein glycosylation [34]. The most abundant proteoform is apoC-III containing one N -acetyl neuraminic acid (NeuAc; sialic acid) attached to a glycan core of one galactose (Gal) and one N -acetylgalactosamine (GalNAc): apoC-III-(Gal) 1 (GalNAc) 1 (NeuAc) 1 .…”

Section: Resultsmentioning

confidence: 99%

Development of multiplex mass spectrometric immunoassay for detection and quantification of apolipoproteins C-I, C-II, C-III and their proteoforms

Trenchevska

Schaab

Nelson

et al. 2015

Methods

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The impetus for discovery and evaluation of protein biomarkers has been accelerated by recent development of advanced technologies for rapid and broad proteome analyses. Mass spectrometry (MS)-based protein assays hold great potential for in vitro biomarker studies. Described here is the development of a multiplex Mass Spectrometric Immunoassay (MSIA) for quantification of apolipoprotein C-I (apoC-I), apolipoprotein C-II (apoC-II), apolipoprotein C-III (apoC-III) and their proteoforms. The multiplex MSIA assay was fast (~40 min) and high-throughput (96 samples at a time). The assay was applied to a small cohort of human plasma samples, revealing the existence of multiple proteoforms for each apolipoprotein C. The quantitative aspect of the assay enabled determination of the concentration for each proteoform individually. Low-abundance proteoforms, such as fucosylated apoC-III, were detected in less than 20% of the samples. The distribution of apoC-III proteoforms varied among samples with similar total apoC-III concentrations. The multiplex analysis of the three apolipoproteins C and their proteoforms using quantitative MSIA represents a significant step forward toward better understanding of their physiological roles in health and disease.

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Mass spectrometry of apolipoprotein C-III, a simple analytical method for mucin-type O-glycosylation and its application to an autosomal recessive cutis laxa type-2 (ARCL2) patient

Cited by 40 publications

References 39 publications

MALDI‐MS profiling of serum O‐glycosylation and N‐glycosylation in COG5‐CDG

MALDI‐MS profiling of serum O‐glycosylation and N‐glycosylation in COG5‐CDG

LC–MS/MS Identification of the O-Glycosylation and Hydroxylation of Amino Acid Residues of Collagen α-1 (II) chain from Bovine Cartilage

Development of multiplex mass spectrometric immunoassay for detection and quantification of apolipoproteins C-I, C-II, C-III and their proteoforms

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