Mass spectrometric and linear discriminant analysis of N-glycans of human serum alpha-1-acid glycoprotein in cancer patients and healthy individuals

Imre, Tı́mea; Kremmer, T.; Héberger, Károly; Molnár-Szöllősi, Éva; Ludányi, Krisztina; Pócsfalvi, Gabriella; Malorni, Antonio; Drahos, László; Vékey, Károly

doi:10.1016/j.jprot.2008.04.005

Cited by 64 publications

(63 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The core fucosylation status of serum haptoglobin glycoforms can be used to differentiate between pancreatic cancer and chronic pancreatitis (43). Serum ␣(1)-acid glycoprotein N-glycosylation patterns in conjunction with linear discriminant analysis were recently used to differentiate between normal, lymphoma, and ovarian cancer cases (44).…”

Section: Discussionmentioning

confidence: 99%

Differential N-glycosylation of Kallikrein 6 Derived from Ovarian Cancer Cells or the Central Nervous System

Kuzmanov

Jiang

Smith

et al. 2009

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Ovarian cancer causes more deaths than any other gynecological disorder. Perturbed glycosylation is one of the hallmarks of this malignancy. Kallikrein 6 (KLK6) elevation in serum is a diagnostic and prognostic indicator in ovarian cancer. The majority of ovarian carcinomas express high levels of KLK6, which diffuses into the circulation. Under physiological conditions, KLK6 is expressed highly in the central nervous system and found at high levels in cerebrospinal fluid from where it enters the circulation. Our aim was to characterize and compare the N-glycosylation status of this protein in ovarian cancer ascites fluid and cerebrospinal fluid. Anion-exchange chromatography was used to reveal different post-translational modifications on the two isoforms. Mobility gel shift Western blot analysis coupled with glycosidase digestion showed that the molecular weight difference between the two isoforms was because of differential glycosylation patterns. The presence of a single N-glycosylation site on KLK6 was confirmed by site-directed mutagenesis. Using a Sambucus nigra agglutinin-monoclonal antibody sandwich enzymelinked immunosorbent assay approach, it was shown that ovarian cancer-derived KLK6 was modified with ␣2-6-linked sialic acid. The structure and composition of glycans of both KLK6 isoforms was elucidated by glycopeptide monitoring with electrospray ionization-Orbitrap tandem mass spectrometry. Therefore, the extensive and almost exclusive sialylation of KLK6 from ovarian cancer cells could lead to the development of an improved biomarker for the early diagnosis of ovarian carcinoma. Molecular & Cellular Proteomics 8:791-798, 2009.

show abstract

Section: Discussionmentioning

confidence: 99%

Differential N-glycosylation of Kallikrein 6 Derived from Ovarian Cancer Cells or the Central Nervous System

Kuzmanov

Jiang

Smith

et al. 2009

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…In order to study glycoforms, the first step nearly always is enzymatic digestion. Often the sugar chain is cleaved off the peptide backbone, resulting in an oligosaccharide mixture [6][7][8][9]]. An alternative is to cleave the peptide chain using proteolytic enzymes (usually trypsin), which results in a peptide/glycopeptide mixture.…”

Section: ) Introductionmentioning

confidence: 99%

Fragmentation characteristics of glycopeptides

Vékey

Ozohanics

Tóth

et al. 2013

International Journal of Mass Spectrometry

Self Cite

View full text Add to dashboard Cite

Mass spectrometric analysis of glycopeptides is an emerging strategy for analysis of glycosylation patterns. Here we present an approach using energy resolved collision induced decomposition (CID) spectra to determine structural features of glycopeptides. Fragmentation of multiply protonated glycopeptides proceeds by a series of competing charge separation processes by cleavage of a glycosidic bond, each producing two charged products: a singly charged, "B" type sugar (oxonium) ion, and a complementary high mass fragment. Energy requirements (activation energies) of these processes are similar to each other, and are far less, than that required for peptide fragmentation. At higher collision energies these first generation products fragment further, yielding a complex fragmentation pattern. Analysis of low energy spectra (those corresponding to ca. 50% survival yield) are straightforward; the ions observed correspond to structural features present in the oligosaccharide, and are not complicated by consecutive reactions. This makes it feasible to identify and distinguish antenna-and core-fucosylated isomers; antenna fucosylation usually suggests presence of the Lewis-X antigen. In general, analysis of the triply protonated molecules are most advantageous, where neutral losses and monosaccharide oxonium ion formation are less abundant.

show abstract

“…Another example is creatine kinase B, which has been found to be upregulated in ovarian cancer cells in vitro and in vivo, and its enzyme activity is significantly elevated in sera from ovarian cancer patients (Huddleston et al, 2005). We also identified potential low-abundance biomarkers such as alpha-1 acid glycoprotein (Imre et al, 2008;Budai et al, 2009) (average PSM of 4.5), cysteine endopeptidases calpain (average PSM of 0.5) (Salehin et al, 2012), SPARC ( Mok et al, 1996, Said et al, 2008Chen et al, 2012) (average PSM of 1.5), tetranectin (average PSM of 8.17) (Hogdall et al, 1993, Hogdall et al, 1994Deng et al, 2000;Begum et al, 2009Begum et al, , 2010, and periostin (Zhu et al, 2010;Morra and Moch, 2011) (average PSM of 4.5) in chicken serum, and peroxiredoxin-1 (average PSM of 2) (Hoskins et al, 2011) in chicken plasma in this study. Therefore, our dataset provides many potential ovarian cancer biomarkers that await validation using MRM-based targeted proteomics because no antibodies to these proteins are available.…”

Section: Discussionmentioning

confidence: 99%

Towards an Animal Model of Ovarian Cancer: Cataloging Chicken Blood Proteins Using Combinatorial Peptide Ligand Libraries Coupled with Shotgun Proteomic Analysis for Translational Research

Sun²,

Matos³

et al. 2014

OMICS: A Journal of Integrative Biology

View full text Add to dashboard Cite

Epithelial ovarian cancer is the most deadly gynecological cancer around the world, with high morbidity in industrialized countries. Early diagnosis is key in reducing its morbidity rate. Yet, robust biomarkers, diagnostics, and animal models are still limited for ovarian cancer. This calls for broader omics and systems science oriented diagnostics strategies. In this vein, the domestic chicken has been used as an ovarian cancer animal model, owing to its high rate of developing spontaneous epithelial ovarian tumors. Chicken blood has thus been considered a surrogate reservoir from which cancer biomarkers can be identified. However, the presence of highly abundant proteins in chicken blood has compromised the applicability of proteomics tools to study chicken blood owing to a lack of immunodepletion methods. Here, we demonstrate that a combinatorial peptide ligand library (CPLL) can efficiently remove highly abundant proteins from chicken blood samples, consequently doubling the number of identified proteins. Using an integrated CPLL-1DGE-LC-MSMS workflow, we identified a catalog of 264 unique proteins. Functional analyses further suggested that most proteins were coagulation and complement factors, blood transport and binding proteins, immune-and defense-related proteins, proteases, protease inhibitors, cellular enzymes, or cell structure and adhesion proteins. Semiquantitative spectral counting analysis identified 10 potential biomarkers from the present chicken ovarian cancer model. Additionally, many human homologs of chicken blood proteins we have identified have been independently suggested as diagnostic biomarkers for ovarian cancer, further triangulating our novel observations reported here. In conclusion, the CPLL-assisted proteomic workflow using the chicken ovarian cancer model provides a feasible platform for translational research to identify ovarian cancer biomarkers and understand ovarian cancer biology. To the best of our knowledge, we report here the most comprehensive survey of the chicken blood proteome to date.

show abstract

Mass spectrometric and linear discriminant analysis of N-glycans of human serum alpha-1-acid glycoprotein in cancer patients and healthy individuals

Cited by 64 publications

References 47 publications

Differential N-glycosylation of Kallikrein 6 Derived from Ovarian Cancer Cells or the Central Nervous System

Differential N-glycosylation of Kallikrein 6 Derived from Ovarian Cancer Cells or the Central Nervous System

Fragmentation characteristics of glycopeptides

Towards an Animal Model of Ovarian Cancer: Cataloging Chicken Blood Proteins Using Combinatorial Peptide Ligand Libraries Coupled with Shotgun Proteomic Analysis for Translational Research

Contact Info

Product

Resources

About