Differential proteomic profile of X- and Y- sorted Sahiwal bull semen

The identification of differential proteins between X- and Y-sperm may be useful for immunological sexing of sperm. Hence, the present study was aimed to compare the protein profile of X- and Y-sorted Sahiwal bull semen using SDS-PAGE and Liquid Chromatography coupled with Mass Spectrometry (Nano LC-MS). Semen sample (n = 6) were categorized into three groups i.e., group I (X-sorted), group II (Y-sorted) and control group (both X- and Y- sperms). SDS PAGE revealed specific proteins of molecular weight between 18 and 24 kDa and between 30 and 37 kDa were present in X-sorted sperms. Also, band corresponding to 25 kDa was specific to Y-sorted sperms. Data obtained from Nano LC/MS is analysed by search engine database i.e., MASCOT and SEQUEST HT. Total, 241 proteins were identified, out of which 113 were differentially expressed between X- and Y-sorted sperms, in which 54 proteins showed at least two unique peptides. Out of 54 proteins, 27 were upregulated in X-sorted sample, 3 were upregulated in Y-sorted sample and 24 were differentially downregulated. Highly upregulated protein in X-sperm viz. Armadillo repeat containing 12 protein, NDC1 transmembrane nucleoporin, β-nerve growth factor, C-type natriuretic peptide, Nucleobindin-2, Phosphoglycerate mutase 2, Calmodulin along with one uncharacterised protein having accession number F1MN9 may have potential to be used as biomarker for separating X and Y sperm.