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BOR - Papers in Press, published online ahead of print June 30, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.031567
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Submitted May 4, 2004
Returned for revision June 3, 2004
Accepted June 21, 2004

Male Reproductive Tract


Antimicrobial Activity of Human Eppin, an Androgen Regulated Sperm Bound Protein with a Whey Acidic Protein Motif

Suresh Yenugu , Richard T. Richardson , Perumal Sivashanmugam , Zengjun Wang , Michael G. O'Rand , Frank S. French , and Susan H. Hall *

* To whom correspondence should be addressed. E-mail: shh{at}med.unc.edu.

Abstract
The role of epididymal sperm binding proteins in reproductive tract immunity is now well recognized in addition to their role in sperm maturation. Sperm acquire forward motility and fertilizing ability during their passage through the epididymis where they acquire a wide variety of proteins belonging to different classes. Previously we demonstrated that Eppin, an androgen regulated sperm binding protein containing protease inhibitory motifs is expressed specifically in the testis and epididymis. In this study we investigated the antibacterial activity of Eppin against E. coli and the mechanism of antimicrobial action. Eppin exhibited dose and time dependent antibacterial activity that was relatively salt insensitive. However, Eppin lost its antibacterial activity completely upon reduction and alkylation of its cysteines indicating the importance of disulfide bonds for its activity. Eppin permeabilized the outer and inner membranes of E. coli consistent with its ability to induce striking morphological alterations of E. coli membranes as shown by scanning electron microscopy. Eppin did not cause disruption of eukaryotic membranes in the rat erythrocyte hemolytic assay. Results of this study indicate that Eppin has a role in the innate immune system of human epididymis.

Key words: Immunology • Male Reproductive Tract • Epididymis • Sperm maturation • Spermatogenesis


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