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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Bioinformatic prediction of the structure and characteristics of human sperm acrosome membrane-associated protein 1 (hSAMP32) and evaluation of its antifertility function in vivo

Tianwu Zhang A * , Junmin Wang B * , Wenbin Niu C , Fang Wang C , Jin Liu B , Yinpei Xing B , Peijun Jia B , Xiuhua Ren B , Jiarui Wang D , Weidong Zang B and Xuemei Chen https://orcid.org/0000-0001-5614-6859 B E
+ Author Affiliations
- Author Affiliations

A School of Computer Science, Henan University of Engineering, Xinzheng, Henan Province, 451191, China.

B Department of Human Anatomy, Basic Medical College of Zhengzhou University, Zhengzhou, Henan Province, 450001, China.

C Reproductive Medical Center of the First Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan Province, 450052, China.

D School of Public Health, Johns Hopkins University, Baltimore, MD 21218, USA.

E Corresponding author. Email: xxchxm@163.com

Reproduction, Fertility and Development 32(16) 1282-1292 https://doi.org/10.1071/RD20198
Submitted: 4 August 2020  Accepted: 8 October 2020   Published: 23 November 2020

Abstract

Human sperm acrosome membrane-associated protein 1 (hSAMP32) plays an important role in the acrosome reaction, sperm–egg primary binding, secondary binding and fusion processes. However, its spatial structural and in vivo antifertility function remain unknown. In this study, we first analysed the physical and chemical characteristics and antigenic epitopes of immunised mice using bioinformatics. Then, we constructed the prokaryotic expression vector pcDNA3.1-hSAMP32 to immunise BALB/c mice in vivo. IgG antibodies in the serum were detected, and the litter size of female mice and the number of the hamster eggs penetrated were counted. hSAMP32 was found to contain six hydrophilic regions and a signal peptide beginning at amino acid position 29. The transmembrane region of hSAMP32 was located within amino acids 217–239 with α-helices and random coil structures. We predicted five antigenic epitopes. The molecular weight of hSAMP32 was 59 kDa. Moreover, the results of in vivo studies revealed that 56 days after the first immunisation, the litter size was significantly smaller for female pcDNA-3.1(+)-hSAMP32-immunised (mean ± s.d. 4.33 ± 1.21) than control mice (9.50 ± 0.55), indicating that the immunocontraception vaccine had an antifertility effect. This experiment presents a theoretical and experimental basis for in-depth study of the hSAMP32 mechanism within the sperm-egg fusing process and for the screening of antigenic epitopes with immunocontraceptive properties.

Graphical Abstract Image

Keywords: bioinformatics, Homo sapiens sperm acrosome associated 1 (hSPACA1), human sperm acrosome membrane-associated protein 1 (hSAMP32), immune infertility, immunocontraception, protein expression, protein structure.


References

Agadjanyan, M. G., Ghochikyan, A., Petrushina, I., Vasilevko, V., Movsesyan, N., Mkrtichyan, M., Saing, T., and Cribbs, D. H. (2005). Prototype Alzheimer’s disease vaccine using the immunodominant B cell epitope from beta-amyloid and promiscuous T cell epitope pan HLA DR-binding peptide. J. Immunol. 174, 1580–1586.
Prototype Alzheimer’s disease vaccine using the immunodominant B cell epitope from beta-amyloid and promiscuous T cell epitope pan HLA DR-binding peptide.Crossref | GoogleScholarGoogle Scholar | 15661919PubMed |

Berger, E., Soldati, R., Huebener, N., Hohn, O., Stermann, A., Durmus, T., Lobitz, S., Zenclussen, A. C., Christiansen, H., Lode, H. N., and Fest, S. (2013). Salmonella SL7207 application is the most effective DNA vaccine delivery method for successful tumor eradication in a murine model for neuroblastoma. Cancer Lett. 331, 167–173.
Salmonella SL7207 application is the most effective DNA vaccine delivery method for successful tumor eradication in a murine model for neuroblastoma.Crossref | GoogleScholarGoogle Scholar | 23337288PubMed |

Chen, Y., Zhang, D., Xin, N., Xiong, Y., Chen, P., Li, B., Tu, X., and Lan, F. (2008). Construction of sperm-specific lactate dehydrogenase DNA vaccine and experimental study of its immunocontraceptive effect on mice. Sci. China C Life Sci. 51, 308–316.
Construction of sperm-specific lactate dehydrogenase DNA vaccine and experimental study of its immunocontraceptive effect on mice.Crossref | GoogleScholarGoogle Scholar | 18368308PubMed |

Chen, X., Liu, X., Ren, X., Li, X., Wang, L., and Zang, W. (2016). Discovery of human posterior head 20 (hPH20) and Homo sapiens sperm acrosome associated 1 (hSPACA1) immunocontraceptive epitopes and their effects on fertility in male and female mice. Reprod. Fertil. Dev. 28, 416–427.
Discovery of human posterior head 20 (hPH20) and Homo sapiens sperm acrosome associated 1 (hSPACA1) immunocontraceptive epitopes and their effects on fertility in male and female mice.Crossref | GoogleScholarGoogle Scholar | 25209425PubMed |

Domagala, A., and Kurpisz, M. (2004). Identification of sperm immunoreactive antigens for immunocontraceptive purposes: a review. Reprod. Biol. Endocrinol. 2, 11.
Identification of sperm immunoreactive antigens for immunocontraceptive purposes: a review.Crossref | GoogleScholarGoogle Scholar | 15035665PubMed |

Fujihara, Y., Satouh, Y., Inoue, N., Isotani, A., Ikawa, M., and Okabe, M. (2012). SPACA1-deficient male mice are infertile with abnormally shaped sperm heads reminiscent of globozoospermia. Development 139, 3583–3589.
SPACA1-deficient male mice are infertile with abnormally shaped sperm heads reminiscent of globozoospermia.Crossref | GoogleScholarGoogle Scholar | 22949614PubMed |

Han, X. N., Li, Q., Lan, X., El-Mufti, L., Ren, H., and Wang, J. (2019). Microglial depletion with clodronate liposomes increases proinflammatory cytokine levels, induces astrocyte activation, and damages blood vessel integrity. Mol. Neurobiol. 56, 6184–6196.
Microglial depletion with clodronate liposomes increases proinflammatory cytokine levels, induces astrocyte activation, and damages blood vessel integrity.Crossref | GoogleScholarGoogle Scholar |

Hao, Z., Wolkowicz, M. J., Shetty, J., Klotz, K., Bolling, L., Sen, B., Westbrook, V. A., Coonrod, S., Flickinger, C. J., and Herr, J. C. (2002). SAMP32, a testis-specific, isoantigenic sperm acrosomal membrane-associated protein. Biol.Reprod. 66, 735–744.
SAMP32, a testis-specific, isoantigenic sperm acrosomal membrane-associated protein.Crossref | GoogleScholarGoogle Scholar | 11870081PubMed |

Jones, R., James, P., and Oxley, D. (2008). The equatorial subsegment in mammalian spermatozoa is enriched in tyrosine phosphorylated proteins. Biol. Reprod. 79, 421–431.
The equatorial subsegment in mammalian spermatozoa is enriched in tyrosine phosphorylated proteins.Crossref | GoogleScholarGoogle Scholar | 18448843PubMed |

Kaur, K., and Prabha, V. (2014). Immunocontraceptives: new approaches to fertility control. BioMed Res. Int. 2014, 868196.
Immunocontraceptives: new approaches to fertility control.Crossref | GoogleScholarGoogle Scholar | 25110702PubMed |

Kawaguchi, A., Momose, F., and Nagata, K. (2011). Replication-coupled and host factor-mediated encapsidation of the influenza virus genome by viral nucleoprotein. J. Virol. 85, 6197–6204.
Replication-coupled and host factor-mediated encapsidation of the influenza virus genome by viral nucleoprotein.Crossref | GoogleScholarGoogle Scholar | 21507964PubMed |

Kishida, K., Harayama, H., Kimura, F., and Murakami, T. (2016). Individual differences in the distribution of sperm acrosome-associated 1 proteins among male patients of infertile couples; their possible impact on outcomes of conventional in vitro fertilization. Zygote 24, 654–661.
Individual differences in the distribution of sperm acrosome-associated 1 proteins among male patients of infertile couples; their possible impact on outcomes of conventional in vitro fertilization.Crossref | GoogleScholarGoogle Scholar | 27185107PubMed |

Luo, J., Yang, J., Cheng, Y., Li, W., Yin, T. L., Xu, W. M., and Zou, Y. J. (2012). Immunogenicity study of plasmid DNA encoding mouse cysteine-rich secretory protein-1 (mCRISP1) as a contraceptive vaccine. Am. J. Reprod. Immunol. 68, 47–55.
Immunogenicity study of plasmid DNA encoding mouse cysteine-rich secretory protein-1 (mCRISP1) as a contraceptive vaccine.Crossref | GoogleScholarGoogle Scholar | 22429321PubMed |

Naz, R. K. (2006). Effect of fertilization antigen (FA-1) DNA vaccine on fertility of female mice. Mol.Reprod. Dev. 73, 1473–1479.
Effect of fertilization antigen (FA-1) DNA vaccine on fertility of female mice.Crossref | GoogleScholarGoogle Scholar | 16894551PubMed |

Naz, R. K. (2011). Antisperm contraceptive vaccines: where we are and where we are going? Am. J. Reprod. Immunol. 66, 5–12.
Antisperm contraceptive vaccines: where we are and where we are going?Crossref | GoogleScholarGoogle Scholar | 21481057PubMed |

Naz, R. K. (2014). Vaccine for human contraception targeting sperm Izumo protein and YLP12 dodecamerpeptide. Protein Sci. 23, 857–868.
Vaccine for human contraception targeting sperm Izumo protein and YLP12 dodecamerpeptide.Crossref | GoogleScholarGoogle Scholar | 24723387PubMed |

Naz, R. K., Gupta, S. K., Gupta, J. C., Vyas, H. K., and Talwar, A. G. (2005). Recent advances in contraceptive vaccine development: a mini-review. Hum. Reprod. 20, 3271–3283.
Recent advances in contraceptive vaccine development: a mini-review.Crossref | GoogleScholarGoogle Scholar | 16113040PubMed |

Ogura, Y., Takagishi, Y., and Harayama, H. (2016). Changes in the distribution and molecular mass of boar sperm acrosome-associated 1 proteins during the acrosome reaction; their validity as indicators for occurrence of the true acrosome reaction. Anim. Reprod. Sci. 172, 94–104.
Changes in the distribution and molecular mass of boar sperm acrosome-associated 1 proteins during the acrosome reaction; their validity as indicators for occurrence of the true acrosome reaction.Crossref | GoogleScholarGoogle Scholar | 27449406PubMed |

Petersen, T. N., Brunak, S., von Heijne, G., and Nielsen, H. (2011). SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat. Methods 8, 785–786.
SignalP 4.0: discriminating signal peptides from transmembrane regions.Crossref | GoogleScholarGoogle Scholar | 21959131PubMed |

Qu, B., Rosenberg, R. N, Li, L., Boyer, P. J., and Johnston, S. A. (2004). Gene vaccination to bias the immune response to amyloid-beta peptide as therapy for Alzheimer disease. Arch. Neurol. 61, 1859–1864.
Gene vaccination to bias the immune response to amyloid-beta peptide as therapy for Alzheimer disease.Crossref | GoogleScholarGoogle Scholar | 15596606PubMed |

Shen, Z. G., He, W., Zhang, J., He, H. Y., Yang, X., Chen, Z. Q., Yang, P., Li, J., Liang, Z. Q., Wu, Y. Z., and Li, J. T. (2011). Induction of specific immune response and suppression of fertility by B-cell-epitope-based mimovirus vaccine. Reproduction 142, 659–666.
Induction of specific immune response and suppression of fertility by B-cell-epitope-based mimovirus vaccine.Crossref | GoogleScholarGoogle Scholar | 21908656PubMed |

Speidel, J. J., Weiss, D. C., Ethelston, S. A., and Gilbert, S. M. (2009). Population policies, programmes and the environment. Philos. Trans. R. Soc. Lond. B Biol. Sci. 364, 3049–3065.
Population policies, programmes and the environment.Crossref | GoogleScholarGoogle Scholar | 19770155PubMed |

Sun, H., Luo, H., Wang, N., Ou, X., Wen, X., Jiang, Y., and Mei, Z. (2013). The expression and purification of TRPM2 protein in E.coli and preparation of its polyclonal antibody. Xibao Yu FenziMianyixue Zazhi 29, 511–514.

Sungsuwan, S., Jongkaewwattana, A., and Jaru-Ampornpan, P. (2020). Nucleocapsid proteins from other swine enteric coronaviruses differentially modulate PEDV replication. Virology 540, 45–56.
Nucleocapsid proteins from other swine enteric coronaviruses differentially modulate PEDV replication.Crossref | GoogleScholarGoogle Scholar | 31756532PubMed |

Suri, A. (2005). Contraceptive vaccines targeting sperm. Expert Opin. Biol. Ther. 5, 381–392.
Contraceptive vaccines targeting sperm.Crossref | GoogleScholarGoogle Scholar | 15833075PubMed |

Tung, K. S., Primakoff, P., Woolman-Gamer, L., and Myles, D. G. (1997). Mechanism of infertility in male guinea pigs immunized with sperm PH-20. Biol. Reprod. 56, 1133–1141.
Mechanism of infertility in male guinea pigs immunized with sperm PH-20.Crossref | GoogleScholarGoogle Scholar | 9160711PubMed |

World Health Organization (WHO) 2010. WHO laboratory manual for the examination and processing of human semen, 5th edition. Geneva: WHO.

Yoon, Y. E., Kim, T. Y., Shin, T. E., Lee, E., Choi, K. H., Lee, S. R., Hong, Y. K., Park, D. S., and Kim, D. K. (2020). Validation of SwimCountTM, a novel home-based device that detects progressively motile spermatozoa: correlation with World Health Organization 5th semen analysis. World J. Mens Health 38, 191–197.
Validation of SwimCountTM, a novel home-based device that detects progressively motile spermatozoa: correlation with World Health Organization 5th semen analysis.Crossref | GoogleScholarGoogle Scholar | 30799559PubMed |

Zelenin, A. V., Kolesnikov, V. A., Tarasenko, O. A., Shafei, R. A., Zelenina, I. A., Mikhailov, V. V., Semenova, M. L., Kovalenko, D. V., Artemyeva, O. V., Ivaschenko, T. E., Evgrafov, O. V., Dickson, G., and Baranovand, V. S. (1997). Bacterial β-galactosidase and human dystrophin genes are expressed in mouse skeletal muscle fibers after ballistic transfection. FEBS Lett. 414, 319–322.
Bacterial β-galactosidase and human dystrophin genes are expressed in mouse skeletal muscle fibers after ballistic transfection.Crossref | GoogleScholarGoogle Scholar | 9315710PubMed |