Subsellular location and characterization of sperm thiol proteins

CABRILLANA M.E; MONCLUS MA; VINCENTI A.E; FORNÉS M.W; YUNES R

Villa Carlos Paz, Córdoba. Argentina

Congreso; XLIV Reunión Anual-Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2008

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB)

Subsellular location and characterization of sperm thiol proteins.  Cabrillana ME, Monclus MA, Vincenti AE, Fornés MW and Yunes R.      During epididymal trip the thiol proteins support a thiol oxidation. This redox changes from thiols to disulfide bonds are associating with the acquisition of sperm motility, capacitation, acrosome reaction and fertilization. Ours objectives were isolate different sperm sub cellular fractions with proteins rich in thiol groups. Using different detergents and sucrose gradient, we fractioned this cell such as head, acrosomal proteins, principal and terminal piece, outer dense fibbers and fibrous sheath. Then we certified the success of this procedure by electron microscopy. On the other hand, we used monobromobimane, a fluorescent radioactive that labels with proteins thiols group. We localized, by fluorescence microscopy, which cellular fraction reacted. Others aliquots were prepared for SDS-PAGE. About 27 KDa band displays a great fluorescence as well as the tail of the sperm. With the positive band we raised polyclonal antibodies     immunizing rabbits. We saw an immune positive reaction with sperm tails. These results suggest that successfully isolate proteins/s rich in thiols groups and we are in condition to initiates the molecular characterization of this protein/s.