Immunological characterization of a mucin-associated protein from hamster tracheal epithelial cell culture.
Airway mucins are high molecular mass (>10(6) dalton) glycoproteins with various types of associated molecules including glycoproteins, lipoproteins, and lipids. The study of mucin-associated proteins is limited largely due to the lack of specific probes. In this study, we produced a monoclonal antibody, MAbHT10, against a 190-kDa mucin associated-protein by immunizing mice with hamster airway mucin purified in nondissociative condition. Using HT10, the 190-kDa mucin-associated protein was characterized immunologically. The 190-kDa mucin-associated protein is glycoprotein and HT10 recognized carbohydrate containing portion of the protein. The association of 190-kDa protein with mucin is strong enough that heat and detergent treatment is required to dissociate it from mucin as evidenced by gel filtration chromatography, Western blot, enzyme-linked immunoadsorbent assay (ELISA), and co-immunoprecipitation. The expression of the 190-kDa protein is increased with the development of hamster tracheal epithelial cells in culture, but showed differences with the pattern of the regulation of mucin expression. Adenosine triphosphate (ATP), a known strong mucin secretagogue, dose-dependently increased mucin release but caused only marginal increase in the release of the 190-kDa protein. The MAb should be useful in the structural and functional analysis of the 190-kDa mucin-associated proteins in physiological and pathological situations such as chronic airway diseases.
Park
Y
Y
Laboratory of Pharmacology, College of Pharmacy, Seoul National University, Korea.
Shin
C Y
CY
Lee
W J
WJ
Jo
M J
MJ
Ryu
J R
JR
Choi
E Y
EY
Kim
K C
KC
Ko
K H
KH
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D018929:Cell Culture Techniques; D006224:Cricetinae; D003902:Detergents / Q000494:pharmacology; D004847:Epithelial Cells / Q000737:chemistry* / Q000166:cytology; D000939:Epitopes / Q000032:analysis; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000276:immunology* / Q000378:metabolism*; D006358:Hot Temperature; D051379:Mice; D008970:Molecular Weight; D009077:Mucins / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding / Q000187:drug effects; D014132:Trachea / Q000737:chemistry* / Q000166:cytology
D000911:Antibodies, Monoclonal; D003902:Detergents; D000939:Epitopes; D006023:Glycoproteins; D009077:Mucins; D000255:Adenosine Triphosphate