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Proteomics. 2016 Dec;16(23):2962-2966. doi: 10.1002/pmic.201600315. Epub 2016 Nov 9.

Exploring the oviductal fluid proteome by a lectin-based affinity approach.

Author information

1
Institute of Biotechnology in Animal Production, Interuniversity Department for Agrobiotechnology (IFA Tulln), University of Natural Resources and Applied Life Sciences Vienna, Tulln, Austria.
2
Institute of Animal Breeding and Genetics, Department for Biomedical Sciences, University of Veterinary Medicine Vienna, Vienna, Austria.
3
Institute of Molecular Animal Breeding and Biotechnology, Ludwig-Maximilian University, Munich, Germany.
4
Reproduction Centre Wieselburg, University of Veterinary Medicine Vienna, Vienna, Austria.
5
VetCore Facility for Research, University of Veterinary Medicine Vienna, Vienna, Austria.
6
Analytical Chemistry, Department of Chemistry-Biomedical Center and Science for Life Laboratory, Uppsala University, Uppsala, Sweden.

Abstract

The analysis of glycoproteins in body fluids represents a central task in the study of vital processes. Herein, we assessed the combined use of Concanavalin A and Wheat Germ Agglutinin as ligands to fractionate and enrich glycoproteins from oviductal fluid (OF), which is a source of molecules involved in fertilization. First, the selectivity was corroborated by a gel-based approach using glycoprotein staining and enzymatic deglycosylation. Nanoliquid chromatography-tandem mass spectrometry (nLC-ESI-MS/MS) further allowed the reliable identification of 134 nonbound as well as 130 lectin-bound OF proteins. Enrichment analysis revealed that 77% of the annotated proteins in the lectin-bound fraction were known glycoproteins (p-value [FDR] = 1.45E-31). The low variance of the number of peptide spectrum matches for each protein within replicates indicated a consistent reproducibility of the whole workflow (median CV 17.3% for technical replicates and 20.7% for biological replicates). Taken together, this study highlights the applicability of a lectin-based workflow for the comprehensive analysis of OF proteins and gives for the first time an insight into the broad glycoprotein content of OF.

KEYWORDS:

Concanavalin A; Glycoproteins; Oviductal fluid; Technology; Wheat germ agglutinin

PMID:
27678138
DOI:
10.1002/pmic.201600315
[Indexed for MEDLINE]

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