Identification and characterization of high molecular-mass mucin-like glycoproteins in the plasma membrane of airway epithelial cells.
A previous lectin binding study demonstrated the presence of high molecular-mass mucin-like glycoproteins (HMGP) on the surface of hamster tracheal surface epithelial (HTSE) secretory cells (Proc. Natl. Acad. Sci. USA 1987;84:9304). In the present study, we intended to isolate and characterize these HMGP from the plasma membrane of the primary HTSE cells and then to determine whether or not these membrane HMGP are Muc-1 mucins, a type of mucins originally discovered on the surface of some carcinomas. A subcellular fraction enriched with the plasma membrane was obtained using a sucrose density gradient centrifugation. This fraction contained high molecular-mass glycoconjugates which were excluded from Sepharose CL-4B gel. Biochemical characterization of these glycoconjugates revealed the following characteristics: (1) susceptibility to both pronase and mild alkaline treatments, but totally resistant to proteoglycan-digesting enzymes; (2) partitioning in the detergent phase of Triton X-114 and resistance to digestion by phosphatidylinositol phospholipase C or D; (3) a buoyant density of 1.5 g/ml based on CsCl density gradient centrifugation; (4) polydispersity in terms of both size and charge density; and (5) lack of immunoreactivity with an anti-Muc-1 mucin antibody. We conclude that the plasma membrane of HTSE cells at confluence contains HMGP, which seem to be the integral membrane proteins but different from Muc-1 mucins, and that these membrane HMGP appear to share some similarities with secreted mucins in terms of size and charge.
Paul
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Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, Maryland, USA.
Lee
D I
DI
Hyun
S W
SW
Gendler
S
S
Kim
K C
KC
D000818:Animals; D000906:Antibodies; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002498:Centrifugation; D002586:Cesium; D002712:Chlorides; D002845:Chromatography; D006224:Cricetinae; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells / Q000737:chemistry*; D004983:Ethanolamines; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D008297:Male; D008647:Mesocricetus; D008970:Molecular Weight; D009077:Mucins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D017830:Octoxynol; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010739:Phospholipase D / Q000494:pharmacology; D011092:Polyethylene Glycols; D011233:Precipitin Tests; D011817:Rabbits; D012685:Sepharose; D014132:Trachea / Q000737:chemistry / Q000166:cytology*; D014316:Tritium; D010738:Type C Phospholipases / Q000494:pharmacology
D000906:Antibodies; D002712:Chlorides; D003902:Detergents; D004983:Ethanolamines; D006023:Glycoproteins; D009077:Mucins; D014316:Tritium; D002586:Cesium; D011092:Polyethylene Glycols; D017830:Octoxynol; D012685:Sepharose; C010615:Nonidet P-40; D010738:Type C Phospholipases; D010739:Phospholipase D; C065017:glycoprotein phospholipase D; D043265:Phosphatidylinositol Diacylglycerol-Lyase; C028019:cesium chloride; D019158:N-Acetylneuraminic Acid; C007369:2-diethylaminoethanol