TY - JOUR
T1 - Epitope Mapping of Antidiacylglycerol Kinase α Monoclonal Antibody DaMab-2
AU - Sano, Masato
AU - Kaneko, Mika K.
AU - Kato, Yukinari
N1 - Funding Information:
Y.K. received research funding from Ono Pharmaceutical Co., Ltd. The other authors have no conflict of interest.
Funding Information:
This research was supported, in part, by AMED under grant numbers JP18am0101078 (Y.K.), JP18am0301010 (Y.K.), and JP18ae0101028 (Y.K.), and by JSPS KAKENHI grant number 17K07299 (M.K.K.) and grant number 16K10748 (Y.K.).
Publisher Copyright:
© 2019, Mary Ann Liebert, Inc., publishers.
PY - 2019/2
Y1 - 2019/2
N2 - Diacylglycerol kinase (DGK) is responsible for the enzymatic conversion of diacylglycerol (DG) to phosphatidic acid (PA). Both DG and PA serve as signaling molecules; therefore, DGK functions as a key enzyme between DG- and PA-mediated signaling. DGKα, one of the 10 DGK isozymes, is involved in T cell function and has been shown to localize in the cytoplasm and nucleus. Furthermore, DGKα translocates to the plasma membrane in response to T cell receptor stimulation. Recently, we developed a specific monoclonal antibody (mAb), DaMab-2 (mouse IgG 1 , kappa), against DGKα. DaMab-2 is very useful in immunocytochemical analysis using HeLa cells. In this study, we characterized the binding epitope of DaMab-2 using Western blot and revealed that Cys246, Lys249, Pro252, and Cys253 of DGKα are important for DaMab-2 binding to the DGKα protein. Our findings can be applied for the production of more functional anti-DGKα mAbs.
AB - Diacylglycerol kinase (DGK) is responsible for the enzymatic conversion of diacylglycerol (DG) to phosphatidic acid (PA). Both DG and PA serve as signaling molecules; therefore, DGK functions as a key enzyme between DG- and PA-mediated signaling. DGKα, one of the 10 DGK isozymes, is involved in T cell function and has been shown to localize in the cytoplasm and nucleus. Furthermore, DGKα translocates to the plasma membrane in response to T cell receptor stimulation. Recently, we developed a specific monoclonal antibody (mAb), DaMab-2 (mouse IgG 1 , kappa), against DGKα. DaMab-2 is very useful in immunocytochemical analysis using HeLa cells. In this study, we characterized the binding epitope of DaMab-2 using Western blot and revealed that Cys246, Lys249, Pro252, and Cys253 of DGKα are important for DaMab-2 binding to the DGKα protein. Our findings can be applied for the production of more functional anti-DGKα mAbs.
KW - DGKa
KW - Diacylglycerol kinase
KW - epitope mapping
KW - monoclonal antibody
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U2 - 10.1089/mab.2018.0047
DO - 10.1089/mab.2018.0047
M3 - Article
C2 - 30648914
AN - SCOPUS:85062168876
SN - 2167-9436
VL - 38
SP - 8
EP - 11
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
IS - 1
ER -