Epididymal protease inhibitor (EPPIN) is a protein hub for seminal vesicle-secreted protein SVS2 binding in mouse spermatozoa

Author(s):Show less -

Mariani, Noemia A. P. ^[1] ; Camara, Aline C. ^[1] ; Silva, Alan Andrew S. ^[1] ; Raimundo, Tamiris R. F. ^[1] ; Andrade, Juliana J. ^[1] ; Andrade, Alexandre D. ^[1] ; Rossini, Bruno C. ^{[2, 3]} ; Marino, Celso L. ^{[2, 3]} ; Kushima, Helio ^[1] ; Santos, Lucilene D. ^{[4, 5]} ; Silva, Erick J. R. ^[1]

Affiliation

^[1] Sao Paulo State Univ UNESP, Inst Biosci, Dept Pharmacol, Botucatu, SP - Brazil

^[2] Sao Paulo State Univ UNESP, Biotechnol Inst IBTEC, Botucatu, SP - Brazil

^[3] Sao Paulo State Univ UNESP, Inst Biosci, Dept Genet, Botucatu, SP - Brazil

^[4] Sao Paulo State Univ UNESP, Botucatu Med Sch FMB, Grad Program Trop Dis, Botucatu, SP - Brazil

^[5] Sao Paulo State Univ UNESP, Ctr Study Venoms Venomous Anim CEVAP, Botucatu, SP - Brazil

• Document type: Journal article
• Source: Molecular and Cellular Endocrinology; v. 506, APR 15 2020.
• Web of Science Citations: 0

Abstract

EPPIN is a sperm-surface drug target for male contraception. Here we investigated EPPIN-interacting proteins in mouse spermatozoa. We showed that EPPIN is an androgen-dependent gene, expressed in the testis and epididymis, but also present in the vas deferens, seminal vesicle and adrenal gland. Mature spermatozoa presented EPPIN staining on the head and flagellum. Immunoprecipitation of EPPIN from spermatozoa pre-incubated with seminal vesicle fluid (SVF) followed by LC-MS/MS or Western blot revealed the co-immunoprecipitation of SVS2, SVS3A, SVS5 and SVS6. In silico and Far-Western blot approaches demonstrated that EPPIN binds SVS2 in a protein network with other SVS proteins. Immunofluorescence using spermatozoa pre-incubated with SVF or recombinant SVS2 demonstrated the co-localization of EPPIN and SVS2 both on sperm head and flagellum. Our data show that EPPIN's roles in sperm function are conserved between mouse and human, demonstrating that the mouse is a suitable experimental model for translational studies on EPPIN. (AU)

• FAPESP's process: 17/11363-8 - EPPIN interacting proteins on mouse sperm surface: identification, expression and potential physiological roles
• Grantee: Noemia Aparecida Partelli Mariani
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 16/23025-7 - Characterization of Eppin (Epididymal protease inhibitor) gene expression in mice
• Grantee: Aline Contri Camara
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 15/08227-0 - A study on EPPIN as a drug target for male contraception: developing a new animal model to test the efficacy of male contraceptive drugs
• Grantee: Erick José Ramo da Silva
• Support Opportunities: Research Grants - Young Investigators Grants