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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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

Substances

  • Glycoproteins
  • Proteome
  • Wheat Germ Agglutinins
  • Concanavalin A