University of Veterinary Medicine Vienna - Research portal

Diagrammed Link to Homepage University of Veterinary Medicine, Vienna

Selected Publication:

Publication type: Journal Article
Document type: Short Communication

Year: 2016

Author(s): Yu, H; Reiser, J; Besenfelder, U; Razzazi-Fazeli, E; Bergquist, J; Brem, G; Artemenko, K; Mayrhofer, C

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

Source: Proteomics. 2016; 16(23):2962-2966



Authors Vetmeduni Vienna:

Besenfelder Urban
Brem Gottfried
Itze-Mayrhofer Corina
Razzazi-Fazeli Ebrahim
Yu Hans

Vetmed Research Units
Institute of Animal Breeding and Genetics, Unit of Reproductive Biology
VetCore


Project(s): Identification of molecules that effect spermatozoa and fertilization success


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 Pubmed: Animals
Body Fluids/chemistry*
Concanavalin A/metabolism
Fallopian Tubes/chemistry
Female
Glycoproteins/analysis*
Glycoproteins/metabolism
Male
Proteome/analysis*
Proteome/metabolism
Rabbits
Reproducibility of Results
Spectrometry, Mass, Electrospray Ionization/methods
Wheat Germ Agglutinins/metabolism
Workflow


© University of Veterinary Medicine ViennaHelp and Downloads