Mucin-like glycoprotein secreted by cultured hamster tracheal epithelial cells

Reen Wu, Charles Plopper, P. W. Cheng

Research output: Contribution to journalArticle

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Abstract

We isolated mucin-like glycoproteins from the conditioned medium of primary hamster tracheal epithelial (HTE) cell culture and characterized them biochemically and immunologically. These glycoproteins were purified on Sepharose CL4B after Streptomyces hyaluronidase treatment and then by CsCl-density-gradient centrifugation in the presence of 4 M-guanidinium chloride. The purified glycoproteins were resistant to digestion by chondroitin AC lyase, heparinase, heparitinase and endo-N-acetylglucosaminidases A, D and H, but susceptible to endo β galactosidase and keratanase. SDS/PAGE demonstrated no contamination by low-molecular-mass proteins. The purified glycoproteins showed a peak buoyant density of 1.56 g/ml in CsCl-density-gradient centrifugation, and contained 10% peptide and 90% carbohydrate by weight. Carbohydrates in these glycoproteins contained N-acetylglucosamine, N-acetylgalactosamine, galactose, fucose, sialic acid and a trace amount of mannose, but no uronic acid. Serine and threonine together accounted for 27% of the total amino acid residues. In addition, the mucin-like glycoproteins exhibited blood-group A and B activities, and very strong inhibitory activity for influenza A virus haemagglutination. With the use of the purified glycoprotein as an antigen, six monoclonal antibodies that stained mucus granules in hamster tracheal epithelium were obtained. We characterized the antibody produced by one of the clones, HM D46. We conclude that HTE cells cultured in the serumfree medium secrete a glycoprotein with physicochemical properties similar to those known in various airways mucins.

Original languageEnglish (US)
Pages (from-to)713-718
Number of pages6
JournalBiochemical Journal
Volume277
Issue number3
StatePublished - 1991

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Mucins
Cricetinae
Glycoproteins
Epithelial Cells
Density Gradient Centrifugation
Centrifugation
keratan-sulfate endo-1,4-beta-galactosidase
heparitinsulfate lyase
Chondroitin Lyases
Carbohydrates
Galactosidases
Heparin Lyase
Uronic Acids
Acetylgalactosamine
Acetylglucosaminidase
Hyaluronoglucosaminidase
Fucose
Acetylglucosamine
Guanidine
Influenza A virus

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mucin-like glycoprotein secreted by cultured hamster tracheal epithelial cells. / Wu, Reen; Plopper, Charles; Cheng, P. W.

In: Biochemical Journal, Vol. 277, No. 3, 1991, p. 713-718.

Research output: Contribution to journalArticle

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