Detection of a secreted MUC1/SEC protein by MUC1 isoform specific monoclonal antibodies.
Although MUC1 proteins are known to be secreted by breast cancer cells, the mechanism of their release from the cell is still obscure. Our previously reported MUC1 cDNA sequences suggested the existence of a secreted MUC1 isoform, MUC1/SEC, that includes a sequence of intron 2, and terminates prematurely at a stop codon within this intron. It is thus devoid of a transmembrane domain. As no formal evidence for MUC1/SEC expression at the protein level had been provided, we generated monoclonal antibodies (mAbs) against a peptide sequence (sec peptide) that is unique for the MUC1/SEC protein. Two anti-sec peptide mAbs were obtained which reacted strongly with (a) the immunizing peptide, (b) recombinant MUC1/SEC protein, and (c) MUC1 proteins secreted from breast cancer cells. The immunoreactivity of the anti-sec peptide mAbs with MUC1 proteins secreted by breast cancer cells was specifically inhibited by the sec peptide-it was completely unaffected by a peptide sequence that represents a MUC1 repeat motif. Significantly, the anti-sec peptide mAbs also detected MUC1/SEC protein in sera of breast cancer patients. We have established here that these mAbs recognize the MUC1/SEC isoform via a peptide sequence which is unique for the MUC1/SEC protein. Our studies thus demonstrate that the MUC1/SEC protein is a bona-fide MUC1 isoform and that its expression may contribute to the secretion of MUC1 proteins by secretory epithelial cells in general and breast cancer cells in particular.
Smorodinsky
N
N
Department of Cell Research and Immunology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel.
Weiss
M
M
Hartmann
M L
ML
Baruch
A
A
Harness
E
E
Yaakobovitz
M
M
Keydar
I
I
Wreschner
D H
DH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001943:Breast Neoplasms / Q000737:chemistry; D005260:Female; D006801:Humans; D051379:Mice; D018396:Mucin-1 / Q000032:analysis* / Q000097:blood / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D018396:Mucin-1; D010446:Peptide Fragments; D011994:Recombinant Proteins