SUBCELLULAR LOCATION AND CHARACTERIZATION OF SPERM THIOL PROTEINS

CABRILLANA ME; MONCLUS MA; VINCENTI AE; FORNES MW.; YUNES R.

Córdoba

Congreso; XLIV Reunión Anual SAIB; 2008

SAIB

During epididymal trip the sperm proteins support a thiol oxidation.This redox changes from thiol to disulfides bonds are associatingwith the acquisition of sperm motility, capacitation, acrosomereaction and fertilization. Ours objectives were isolate differentsperm subcellular fractions with proteins rich in thiol groups. Usingdifferent detergent and sucrose gradient, we fractionated this cellsuch as head, acrosomal proteins, principal and terminal piece,outer dense fibers and fibrous sheath. Then we certificated thesuccess of this procedure by electron microscopy. On the other handwe used monobromobimane, a fluorescent reactive that labels withprotein`s thiols groups. We localized, by fluorescence microscopy,witch cellular fraction reacted. Others aliquots, were prepared forSDS-PAGE. About 27 kDa band display a great fluorescence aswell as the tail of the sperm. With this positive band we raisedpolyclonal antibody immunizing rabbits. We saw an immunepositive reaction with sperms tail. These results suggest that wesuccessfully isolated protein/s rich in thiols groups and we are incondition to initiates the molecular characterization of thisprotein/s.