Subcellular location and characterization of sperm thiol proteins

CABRILLANA ME; MONCLUS MA; VINCENTI AE; FORNÉS MW; YUNES R

Villa Carlos Paz Córdoba

Congreso; XLIV Reunión anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2008

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

During epididymal trip the sperm proteins support a thiol oxidation. This redox changes from thiol to disulfides bonds are associating with the acquisition of sperm motility, capacitation, acrosome reaction and fertilization. Ours objectives were isolate different sperm subcellular fractions with proteins rich in thiol groups. Using different detergent and sucrose gradient, we fractionated this cell such as head, acrosomal proteins, principal and terminal piece, outer dense fibers and fibrous sheath. Then we certificated the success of this procedure by electron microscopy. On the other hand we used monobromobimane, a fluorescent reactive that labels with protein`s thiols groups. We localized, by fluorescence microscopy, witch cellular fraction reacted. Others aliquots, were prepared for   SDS-PAGE. About 27 kDa band display a great fluorescence as well as the tail of the sperm. With this positive band we raised polyclonal antibody immunizing rabbits. We saw a positive reaction with sperms tail.  These results suggest that we successfully isolated protein/s rich in thiols groups and we are in condition to initiates the molecular characterization of this protein/s.