Isolation of plasma membrane proteins from bovine spermatozoa using ultracentrifugation

Abstract

In this study, we employed 10% SDS-PAGE to analyze total and plasma membrane proteins from bovine spermatozoa, revealing an average of 17 bands for total sperm proteins and 12 bands for plasma membrane proteins. The molecular weights of the proteins ranged from 21 to 250 kDa in cell lysates and 30 to 250 kDa in plasma membrane proteins. Prominent bands at approximately 26, 50, 60, 67, 100, and 120 kDa were observed in total sperm proteins, while bands at approximately 40, 80, 95, and 110 kDa were noted in plasma membrane proteins. These results highlighting both quantitative and qualitative differences in low molecular mass proteins. The higher molecular weight bands in total sperm proteins may be attributed to variations in protein concentration. Further, 2D electrophoresis identified over 60 protein spots in the sperm membrane, with molecular weights ranging from 37 to 200 kDa. These findings underscore the complexity and variability of sperm protein profiles and suggest that they could improve fertility assessments and semen preservation methods. Future research, including LC-MS analysis and immunoblot techniques, is needed for detailed protein identification and to advance our understanding and application of reproductive technologies.