Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ

Glycoproteins are potentially important biomarkers of disease and therapeutic targets. In particular, the N-linked glycoproteins are a focus of interest as they can be found in the extracellular environment and body fluids. In this study, we have sampled the tears, the extracellular fluid of the epi...

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Autores principales: Zhou, Lei, Beuerman, Roger W., Chew, Ai Ping, Koh, Siew Kwan, Cafaro, Thamara A., Urrets Zavalía, Enrique Alberto, Urrets Zavalía, Julio, Li, Sam F.Y., Serra, Horacio M.
Formato: Artículo
Lenguaje:Español
Publicado: 2009
Materias:
RE Oftalmología
Acceso en línea:http://pa.bibdigital.ucc.edu.ar/4942/1/A_Zhou.pdf
Aporte de:
Producción Académica Universidad Católica de Córdoba (UCCor) de Universidad Católica de Córdoba
id I38-R144-4942
record_format dspace
spelling I38-R144-49422025-10-21T13:48:49Z http://pa.bibdigital.ucc.edu.ar/4942/ Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ Zhou, Lei Beuerman, Roger W. Chew, Ai Ping Koh, Siew Kwan Cafaro, Thamara A. Urrets Zavalía, Enrique Alberto Urrets Zavalía, Julio Li, Sam F.Y. Serra, Horacio M. RE Oftalmología Glycoproteins are potentially important biomarkers of disease and therapeutic targets. In particular, the N-linked glycoproteins are a focus of interest as they can be found in the extracellular environment and body fluids. In this study, we have sampled the tears, the extracellular fluid of the epithelial cells covering the surface of the eye, of patients with climatic droplet keratopathy (CDK) using tears of unaffected normal patients for comparison. Prefractionation of the tear sample used a hydrazide-resin capture method, and the previously N-glycosylated peptides were then subjected to two-dimensional nano-LC-nano-ESI-MS/MS analysis to obtain peptide fragmentation patterns for identification through protein database searches. We have identified a total of 43 unique N-glycoproteins, 19 of which have not previously been reported in tear fluid. In addition, we have quantitatively compared N-glycoprotein profiles in tear fluid of patients with CDK to tears of nondiseased controls using glycopeptide capture, iTRAQ labeling and 2D nano-LC-nano-ESI-MS/MS analysis. In tears of CDK patients, increased levels of four N-glycosylated proteins including haptoglobin (at sites N207, N211 and N241), polymeric immunoglobulin receptor (at sites N83, N90, N135, N186, N421, and N469), immunoglobulin J chain (at site N49) and an uncharacterized protein DKFZp686M08189 (at site N470), as well as a decrease in the N-glycosylation level of one N-glycosylated protein, lacritin (at site N119) were observed. However, the overall levels of these five proteins showed no appreciable changes between control and CDK samples. The findings could be clinically significant in terms of disease etiology and biomarkers. 2009-04-03 info:eu-repo/semantics/article info:eu-repo/semantics/closedAccess application/pdf spa http://pa.bibdigital.ucc.edu.ar/4942/1/A_Zhou.pdf Zhou, Lei, Beuerman, Roger W., Chew, Ai Ping, Koh, Siew Kwan, Cafaro, Thamara A., Urrets Zavalía, Enrique Alberto, Urrets Zavalía, Julio ORCID: https://orcid.org/0000-0003-2267-2783 <https://orcid.org/0000-0003-2267-2783>, Li, Sam F.Y. and Serra, Horacio M. ORCID: https://orcid.org/0000-0003-4979-8642 <https://orcid.org/0000-0003-4979-8642> (2009) Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ. Journal of Proteome Research, 8 (4). pp. 1992-2003. ISSN 19714880 info:eu-repo/semantics/altIdentifier/doi/10.1021/pr800962q
institution Universidad Católica de Córdoba
institution_str I-38
repository_str R-144
collection Producción Académica Universidad Católica de Córdoba (UCCor)
language Español
orig_language_str_mv spa
topic RE Oftalmología
spellingShingle RE Oftalmología
Zhou, Lei
Beuerman, Roger W.
Chew, Ai Ping
Koh, Siew Kwan
Cafaro, Thamara A.
Urrets Zavalía, Enrique Alberto
Urrets Zavalía, Julio
Li, Sam F.Y.
Serra, Horacio M.
Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ
topic_facet RE Oftalmología
description Glycoproteins are potentially important biomarkers of disease and therapeutic targets. In particular, the N-linked glycoproteins are a focus of interest as they can be found in the extracellular environment and body fluids. In this study, we have sampled the tears, the extracellular fluid of the epithelial cells covering the surface of the eye, of patients with climatic droplet keratopathy (CDK) using tears of unaffected normal patients for comparison. Prefractionation of the tear sample used a hydrazide-resin capture method, and the previously N-glycosylated peptides were then subjected to two-dimensional nano-LC-nano-ESI-MS/MS analysis to obtain peptide fragmentation patterns for identification through protein database searches. We have identified a total of 43 unique N-glycoproteins, 19 of which have not previously been reported in tear fluid. In addition, we have quantitatively compared N-glycoprotein profiles in tear fluid of patients with CDK to tears of nondiseased controls using glycopeptide capture, iTRAQ labeling and 2D nano-LC-nano-ESI-MS/MS analysis. In tears of CDK patients, increased levels of four N-glycosylated proteins including haptoglobin (at sites N207, N211 and N241), polymeric immunoglobulin receptor (at sites N83, N90, N135, N186, N421, and N469), immunoglobulin J chain (at site N49) and an uncharacterized protein DKFZp686M08189 (at site N470), as well as a decrease in the N-glycosylation level of one N-glycosylated protein, lacritin (at site N119) were observed. However, the overall levels of these five proteins showed no appreciable changes between control and CDK samples. The findings could be clinically significant in terms of disease etiology and biomarkers.
format Artículo
author Zhou, Lei
Beuerman, Roger W.
Chew, Ai Ping
Koh, Siew Kwan
Cafaro, Thamara A.
Urrets Zavalía, Enrique Alberto
Urrets Zavalía, Julio
Li, Sam F.Y.
Serra, Horacio M.
author_facet Zhou, Lei
Beuerman, Roger W.
Chew, Ai Ping
Koh, Siew Kwan
Cafaro, Thamara A.
Urrets Zavalía, Enrique Alberto
Urrets Zavalía, Julio
Li, Sam F.Y.
Serra, Horacio M.
author_sort Zhou, Lei
title Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ
title_short Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ
title_full Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ
title_fullStr Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ
title_full_unstemmed Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ
title_sort quantitative analysis of n-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and itraq
publishDate 2009
url http://pa.bibdigital.ucc.edu.ar/4942/1/A_Zhou.pdf
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