Identification of Ewing's sarcoma gene product as a glycoprotein using a monoclonal antibody that recognizes an immunodeterminant containing O-linked N-acetylglucosamine moiety.
The Ewing's sarcoma (EWS) oncogene is fused to a variety of cellular transcription factors in various forms of human cancers. Although EWS fusion proteins have been extensively studied, the normal function of EWS remains poorly characterized. We previously reported that a monoclonal antibody, referred to as MY95, recognized nucleoporins such as p62, Nup98, and CAN/Nup214 and an uncharacterized polypeptide with an apparent molecular mass of 83 kDa. In the present study, an amino acid sequence analysis of this 83-kDa protein revealed that it is, in fact, EWS, which is not known to belong to the nucleoporins. We further demonstrated that the immunodeterminant of MY95 contains an N-acetylglucosamine moiety, indicating that EWS is a glycoprotein. Interestingly, the glycosylation level of EWS changes during the neural differentiation of P19 cells. MY95 will be quite useful in further studies of the glycosylated form of EWS in terms of understanding the normal cellular function of this oncogene product.
Department of Cell Biology and Neuroscience, Graduate School of Medicine, Osaka University, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
Yosuke
Y
Matsuoka
Yuriko
Y
Matsuoka
Satoshi
S
Shibata
Noriko
N
Yasuhara
Yoshihiro
Y
Yoneda
D000117:Acetylglucosamine / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D034802:RNA-Binding Protein EWS / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D012512:Sarcoma, Ewing / Q000235:genetics* / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006023:Glycoproteins; D034802:RNA-Binding Protein EWS; D000117:Acetylglucosamine