TY - JOUR
T1 - Identification and characterization of the antigen recognized by the germ cell mAb TRA98 using a human comprehensive wet protein array
AU - Fukuda, Eriko
AU - Tanaka, Hiromitsu
AU - Yamaguchi, Kei
AU - Takasaka, Mieko
AU - Kawamura, Yoshifumi
AU - Okuda, Hidenobu
AU - Isotani, Ayako
AU - Ikawa, Masahito
AU - Shapiro, Virginia Smith
AU - Tsuchida, Junji
AU - Okada, Yuki
AU - Tsujimura, Akira
AU - Miyagawa, Yasushi
AU - Fukuhara, Shinichiro
AU - Kawakami, Yoshitaka
AU - Wada, Morimasa
AU - Nishimune, Yoshitake
AU - Goshima, Naoki
N1 - Publisher Copyright:
© 2021 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd
PY - 2021/3
Y1 - 2021/3
N2 - 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.
AB - 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.
KW - SUMOylation
KW - cell-free protein expression
KW - in vitro proteome
KW - nuclear factor-κB activating protein
KW - protein array
KW - testis
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U2 - 10.1111/gtc.12832
DO - 10.1111/gtc.12832
M3 - Article
C2 - 33527666
AN - SCOPUS:85102152463
SN - 1356-9597
VL - 26
SP - 180
EP - 189
JO - Genes to Cells
JF - Genes to Cells
IS - 3
ER -