Expression of Recombinant BSP3 Protein and its Localization Studies in Buffalo (Bubalus bubalis) Spermatozoa

Autor: A. M. Shende, S. K. Ghosh, Sadhan Bag, V. H. Muhammed Kutty, S.K. Bhure, Harikrishna Pillai
Rok vydání: 2015
Předmět:
Zdroj: Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 87:1067-1072
ISSN: 2250-1746
0369-8211
DOI: 10.1007/s40011-015-0663-1
Popis: The binder sperm proteins (BSP) have damaging effects on the sperm cryopreservation. Greater concentrations of BSPs in seminal plasma remove more cholesterol and phospholipids from the sperm membrane, resulting in a destabilization that decreases sperm resistance to cold shock and freezing. The present study was conducted to clone, express recombinant BSP3 and find its localization in spermatozoa of Bubalus bubalis. The amplified buffalo BSP3 gene coded for 136 amino acids and the expressed recombinant protein showed a molecular weight of about 19.5 kDa. The hyper-immune serum raised against the recombinant BSP3 reacted specifically to a 19.5 kDa protein in the sperm cell lysate. Further, the Bioinformatics analysis predicted two fibronectin type 2 (FN-2) binding domains. The buffalo BSP3 showed changes in the amino acid sequence in both N-terminal and FN2 domains which might alter its binding properties. The protein has been localized to the acrosome and the middle piece of the spermatozoa by indirect immuno-fluorescence microscopy. Nevertheless, the antibodies against BSP3 can be used to measure its concentration in buffalo ejaculates and to effectively sequester excess BSP3 to prevent the early cryocapacitation.
Databáze: OpenAIRE
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