Type of publication:
Type of document:
Yu, H; Reiser, J; Besenfelder, U; Razzazi-Fazeli, E; Bergquist, J; Brem, G; Artemenko, K; Mayrhofer, C
Exploring the oviductal fluid proteome by a lectin-based affinity approach.
Proteomics. 2016; 16(23):2962-2966
Authors Vetmeduni Vienna:
Vetmed Research Units
Institute of Animal Breeding and Genetics, Unit of Reproductive Biology
Identification of molecules that effect spermatozoa and fertilization success
- 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.© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Reproducibility of Results
Spectrometry, Mass, Electrospray Ionizationmethods
Wheat Germ Agglutininsmetabolism