Identification and characterization of high molecular-mass mucin-like glycoproteins in the plasma membrane of airway epithelial cells

Emmanuel Paul, Dong Lee, Sang Won Hyun, Sandra Gendler, K. Chul Kim

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)681-690
Number of pages10
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume19
Issue number4
StatePublished - 1998
Externally publishedYes

Fingerprint

Molecular mass
Mucins
Cell membranes
Glycoproteins
Epithelial Cells
Cell Membrane
Mucin-1
Cricetinae
Glycoconjugates
Density Gradient Centrifugation
Centrifugation
Membranes
Phospholipase D
Pronase
Subcellular Fractions
Membrane Glycoproteins
Polydispersity
Type C Phospholipases
Proteoglycans
Phosphatidylinositols

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

Cite this

Identification and characterization of high molecular-mass mucin-like glycoproteins in the plasma membrane of airway epithelial cells. / Paul, Emmanuel; Lee, Dong; Hyun, Sang Won; Gendler, Sandra; Kim, K. Chul.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 19, No. 4, 1998, p. 681-690.

Research output: Contribution to journalArticle

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