Characterization of a unique mucin-like glycoprotein secreted by a human endometrial adenocarcinoma cell line (Ishikawa).
A human endometrial adenocarcinoma cell line (Ishikawa) has been shown to incorporate [3H]glucosamine and to secrete a radiolabeled high molecular weight compound which is excluded from a Sepharose CL-2B column. The excluded material was resistant to hyaluronidase, chondroitinase ABC, and heparinase. These findings rule out the possibility of this material being a proteoglycan. The susceptibility of this material to digestion with pronase, neuraminidase, and alkaline borohydride treatment strongly suggests that the excluded material is an O-glycosidic glycoprotein. The glycoprotein secreted by Ishikawa cells (ICGP) did not react immunologically with antibodies against either lactoferrin or fibronectin, but did react with an antibody made against tracheal mucin. Conversely, immunoblot analysis revealed that an antibody made against ICGP did not recognize hyaluronic acid, chondroitin, heparin, nasal turbinate mucin, bovine submaxillary gland mucin, lactoferrin, or fibronectin, but did recognize tracheal mucin. Analysis of ICGP amino acid and carbohydrate composition showed that it is rich in serine, threonine, glutamic acid, aspartic acid, and N-acetylneuraminic acid. In this respect, ICGP differs from other mucins, even though it is immunologically similar to respiratory mucin; hence we may consider ICGP to be a mucin-like glycoprotein. Secretion of ICGP can be modulated by Ca(2+)-ionophore and other mucus secretagogues, such as platelet activating factor, carbachol, and monocyte/macrophage mucus secretagogue, all mediators of lung inflammation. Ishikawa cells and anti-ICGP antibody may be used in studies on in vitro regulation of mucin-like glycoprotein synthesis and secretion in the respiratory tract as well as in the endometrium.
Goswami
S
S
Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029.
Gollub
E
E
Weiss
D J
DJ
Gurpide
E
E
Roboz
J
J
Marom
Z
Z
D000230:Adenocarcinoma / Q000737:chemistry / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D000001:Calcimycin / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002241:Carbohydrates / Q000032:analysis; D016889:Endometrial Neoplasms / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D006801:Humans; D007120:Immunochemistry; D009077:Mucins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism
D000596:Amino Acids; D002241:Carbohydrates; D009077:Mucins; D010972:Platelet Activating Factor; D000001:Calcimycin; D002217:Carbachol