Antimicrobial activity of human EPPIN, an androgen-regulated, sperm-bound protein with a whey acidic protein motif.
The role of epididymal sperm-binding proteins in reproductive tract immunity is now well recognized in addition to their role in sperm maturation. Spermatozoa 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 (epididymal protease inhibitor), an androgen-regulated, sperm-binding protein containing protease-inhibitory motifs, is expressed specifically in the testis and epididymis. In the present study, we investigated the antibacterial activity of EPPIN against Escherichia coli and the mechanism of antimicrobial action. EPPIN exhibited dose- and time-dependent antibacterial activity that was relatively insensitive to salt. However, EPPIN lost its antibacterial activity completely on 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, which is 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. The present results indicate that EPPIN has a role in the innate immune system of human epididymis.
Yenugu
Suresh
S
Laboratories for Reproductive Biology, University of North Carolina, Chapel Hill, North Carolina 27599-7500, USA.
Richardson
Richard T
RT
Sivashanmugam
Perumal
P
Wang
Zengjun
Z
O'rand
Michael G
MG
French
Frank S
FS
Hall
Susan H
SH
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000728:Androgens / Q000502:physiology*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006461:Hemolysis; D006801:Humans; D008297:Male; D008855:Microscopy, Electron, Scanning; D008894:Milk Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D051381:Rats; D013094:Spermatozoa / Q000378:metabolism*; D013997:Time Factors
D000728:Androgens; D000900:Anti-Bacterial Agents; C431553:Eppin protein, human; D008894:Milk Proteins; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; C024471:whey acidic proteins