Js. Marken et al., MEMBRANE TOPOLOGY OF THE L6 ANTIGEN AND IDENTIFICATION OF THE PROTEINEPITOPE RECOGNIZED BY THE L6 MONOCLONAL-ANTIBODY, The Journal of biological chemistry, 269(10), 1994, pp. 7397-7401
The murine monoclonal antibody (mAb) L6 recognizes an integral membran
e glycoprotein that is highly expressed on lung, breast, colon, and ov
arian carcinomas and is referred to as the L6 antigen. This antigen is
an attractive target for therapeutic intervention due to its high lev
el expression on malignant cells. We have previously reported the isol
ation of a cDNA encoding the human L6 antigen (H-L6). Here, we report
the isolation of a cDNA clone encoding the murine L6 antigen (M-L6). T
his cDNA contains one long open reading frame, which encodes a 220-ami
no acid polypeptide that is 78% homologous to H-L6. This protein conta
ins short NH2- and COOH-terminal hydrophilic domains and four hydropho
bic regions, each long enough to span the plasma membrane. Each of the
se hydrophobic domains is separated by a hydrophilic domain, the longe
st of which contains one possible N-linked glycosylation site and is l
ocated between the third and fourth hydrophobic domains. We have previ
ously demonstrated that the murine L6 mAb recognizes a protein epitope
expressed on human tumor-derived cell lines. Now, using chimeric cDNA
constructs encoding human-murine L6 antigen hybrids in conjunction wi
th monoclonal antibody binding experiments, we show that the 42-residu
e hydrophilic domain of the L6 antigen, located between the third and
fourth hydrophobic domains, is outside the cell and that residues in t
he NH2-terminal region of this domain are critical for the binding of
the murine L6 mAb to H-L6.