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Exploring the oviductal fluid proteome by a lectin-based affinity approach
- Source :
- PROTEOMICS. 16:2962-2966
- Publication Year :
- 2016
- Publisher :
- Wiley, 2016.
-
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.
- Subjects :
- Male
0301 basic medicine
Spectrometry, Mass, Electrospray Ionization
Proteome
Wheat Germ Agglutinins
Peptide
Biochemistry
Workflow
03 medical and health sciences
Concanavalin A
Animals
Molecular Biology
Fallopian Tubes
Glycoproteins
chemistry.chemical_classification
biology
Reproducibility of Results
Lectin
Molecular biology
Wheat germ agglutinin
Body Fluids
Staining
030104 developmental biology
Enzyme
chemistry
biology.protein
Female
Rabbits
Glycoprotein
Subjects
Details
- ISSN :
- 16159853
- Volume :
- 16
- Database :
- OpenAIRE
- Journal :
- PROTEOMICS
- Accession number :
- edsair.doi.dedup.....97d04394370582583146737ac03a461a