Identification and characterization of the antigen recognized by the germ cell mAb TRA98 using a human comprehensive wet protein array

Eriko Fukuda, Hiromitsu Tanaka, Kei Yamaguchi, Mieko Takasaka, Yoshifumi Kawamura, Hidenobu Okuda, Ayako Isotani, Masahito Ikawa, Virginia Smith Shapiro, Junji Tsuchida, Yuki Okada, Akira Tsujimura, Yasushi Miyagawa, Shinichiro Fukuhara, Yoshitaka Kawakami, Morimasa Wada, Yoshitake Nishimune, Naoki Goshima

Research output: Contribution to journalArticlepeer-review

Abstract

TRA98 is a rat monoclonal antibody (mAb) which recognizes a specific antigen in the nuclei of germ cells. mAb TRA98 has been used to understand the mechanism of germ cell development and differentiation in many studies. In mice, the antigen recognized by mAb TRA98 or GCNA1 has been reported to be a GCNA gene product, but despite the demonstration of the immunoreactivity of this mAb in human testis and sperm in 1997, the antigen in humans remains unknown, as of date. To identify the human antigen recognized by mAb TRA98, a human comprehensive wet protein array was developed containing 19,446 proteins derived from human cDNAs. Using this array, it was found that the antigen of mAb TRA98 is not a GCNA gene product, but nuclear factor-κB activating protein (NKAP). In mice, mAb TRA98 recognized both the GCNA gene product and NKAP. Furthermore, conditional knockout of Nkap in mice revealed a phenotype of Sertoli cell-only syndrome. Although NKAP is a ubiquitously expressed protein, NKAP recognized by mAb TRA98 in mouse testis was SUMOylated. These results suggest that NKAP undergoes modifications, such as SUMOylation in the testis, and plays an important role in spermatogenesis.

Original languageEnglish (US)
Pages (from-to)180-189
Number of pages10
JournalGenes to Cells
Volume26
Issue number3
DOIs
StatePublished - Mar 2021

Keywords

  • cell-free protein expression
  • in vitro proteome
  • nuclear factor-κB activating protein
  • protein array
  • SUMOylation
  • testis

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

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