Abstract
Primary hamster tracheal epithelial cells growing on a collagen gel matrix produce high molecular weight mucins indistinguishable from mucins produced in vivo. Using a modified version of these confluent cultures, we have demonstrated here that (i) release of mucins can be stimulated by human neutrophil elastase (HNE; EC 3.4.21.37); (ii) HNE can degrade mucins, and both mucin release and degradation by HNE require an active catalytic site; and (iii) there are at least two pools of mucins in these cells: one is a rapidly turning-over spontaneously releasable constitutive pool, the other is a slowly turning-over HNE-releasable pool. We provide evidence that the HNE-releasable mucins are membrane bound and associated with the secretory cell apical surface.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adler K. B., Winn W. C., Jr, Alberghini T. V., Craighead J. E. Stimulatory effect of Pseudomonas aeruginosa on mucin secretion by the respiratory epithelium. JAMA. 1983 Mar 25;249(12):1615–1617. [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Breuer R., Lucey E. C., Stone P. J., Christensen T. G., Snider G. L. Proteolytic activity of human neutrophil elastase and porcine pancreatic trypsin causes bronchial secretory cell metaplasia in hamsters. Exp Lung Res. 1985;9(1-2):167–175. doi: 10.3109/01902148509061535. [DOI] [PubMed] [Google Scholar]
- Chandrasekaran E. V., Rana S. S., Davila M., Mendicino J. Structures of the oligosaccharide chains in swine trachea mucin glycoproteins. J Biol Chem. 1984 Oct 25;259(20):12908–12914. [PubMed] [Google Scholar]
- De Water R., Willems L. N., Van Muijen G. N., Franken C., Fransen J. A., Dijkman J. H., Kramps J. A. Ultrastructural localization of bronchial antileukoprotease in central and peripheral human airways by a gold-labeling technique using monoclonal antibodies. Am Rev Respir Dis. 1986 May;133(5):882–890. [PubMed] [Google Scholar]
- Heck L. W., Darby W. L., Hunter F. A., Bhown A., Miller E. J., Bennett J. C. Isolation, characterization, and amino-terminal amino acid sequence analysis of human neutrophil elastase from normal donors. Anal Biochem. 1985 Aug 15;149(1):153–162. doi: 10.1016/0003-2697(85)90488-9. [DOI] [PubMed] [Google Scholar]
- Keenan K. P., Combs J. W., McDowell E. M. Regeneration of hamster tracheal epithelium after mechanical injury. I. Focal lesions: quantitative morphologic study of cell proliferation. Virchows Arch B Cell Pathol Incl Mol Pathol. 1982;41(3):193–214. doi: 10.1007/BF02890281. [DOI] [PubMed] [Google Scholar]
- Kim K. C. Possible requirement of collagen gel substratum for production of mucin-like glycoproteins by primary rabbit tracheal epithelial cells in culture. In Vitro Cell Dev Biol. 1985 Nov;21(11):617–621. doi: 10.1007/BF02623293. [DOI] [PubMed] [Google Scholar]
- Kim K. C., Rearick J. I., Nettesheim P., Jetten A. M. Biochemical characterization of mucous glycoproteins synthesized and secreted by hamster tracheal epithelial cells in primary culture. J Biol Chem. 1985 Apr 10;260(7):4021–4027. [PubMed] [Google Scholar]
- Klinger J. D., Tandler B., Liedtke C. M., Boat T. F. Proteinases of Pseudomonas aeruginosa evoke mucin release by tracheal epithelium. J Clin Invest. 1984 Nov;74(5):1669–1678. doi: 10.1172/JCI111583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marom Z., Shelhamer J. H., Kaliner M. Human pulmonary macrophage-derived mucus secretagogue. J Exp Med. 1984 Mar 1;159(3):844–860. doi: 10.1084/jem.159.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niles R. M., Christensen T. G., Breuer R., Stone P. J., Snider G. L. Serine proteases stimulate mucous glycoprotein release from hamster tracheal ring organ culture. J Lab Clin Med. 1986 Nov;108(5):489–497. [PubMed] [Google Scholar]
- Powers J. C. Reaction of serine proteases with halomethyl ketones. Methods Enzymol. 1977;46:197–208. doi: 10.1016/s0076-6879(77)46020-8. [DOI] [PubMed] [Google Scholar]
- Reid L., Bhaskar K., Coles S. Control and modulation of airway epithelial cells and their secretions. Exp Lung Res. 1983 Feb;4(2):157–170. doi: 10.3109/01902148309055011. [DOI] [PubMed] [Google Scholar]
- Roth J. Cytochemical localization of terminal N-acetyl-D-galactosamine residues in cellular compartments of intestinal goblet cells: implications for the topology of O-glycosylation. J Cell Biol. 1984 Feb;98(2):399–406. doi: 10.1083/jcb.98.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schulte B. A., Spicer S. S. Histochemical methods for characterizing secretory and cell surface sialoglycoconjugates. J Histochem Cytochem. 1985 May;33(5):427–438. doi: 10.1177/33.5.3989272. [DOI] [PubMed] [Google Scholar]
- Senior R. M., Bielefeld D. R., Starcher B. C. Comparison of the elastolytic effects of human leukocyte elastase and porcine pancreatic elastase. Biochem Biophys Res Commun. 1976 Oct 18;72(4):1327–1334. doi: 10.1016/s0006-291x(76)80160-x. [DOI] [PubMed] [Google Scholar]
- Snider G. L., Lucey E. C., Christensen T. G., Stone P. J., Calore J. D., Catanese A., Franzblau C. Emphysema and bronchial secretory cell metaplasia induced in hamsters by human neutrophil products. Am Rev Respir Dis. 1984 Jan;129(1):155–160. doi: 10.1164/arrd.1984.129.1.155. [DOI] [PubMed] [Google Scholar]
- Snider G. L., Stone P. J., Lucey E. C., Breuer R., Calore J. D., Seshadri T., Catanese A., Maschler R., Schnebli H. P. Eglin-c, a polypeptide derived from the medicinal leech, prevents human neutrophil elastase-induced emphysema and bronchial secretory cell metaplasia in the hamster. Am Rev Respir Dis. 1985 Dec;132(6):1155–1161. doi: 10.1164/arrd.1985.132.6.1155. [DOI] [PubMed] [Google Scholar]
- Specian R. D., Neutra M. R. Cytoskeleton of intestinal goblet cells in rabbit and monkey. The theca. Gastroenterology. 1984 Dec;87(6):1313–1325. [PubMed] [Google Scholar]
- Spicer S. S., Martinez J. R. Mucin biosynthesis and secretion in the respiratory tract. Environ Health Perspect. 1984 Apr;55:193–204. doi: 10.1289/ehp.8455193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stone P. J., Calore J. D., Snider G. L., Franzblau C. Role of alpha-macroglobulin-elastase complexes in the pathogenesis of elastase-induced emphysema in hamsters. J Clin Invest. 1982 Apr;69(4):920–931. doi: 10.1172/JCI110531. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stone P. J., Franzblau C., Kagan H. M. Proteolysis of insoluble elastin. Methods Enzymol. 1982;82(Pt A):588–605. doi: 10.1016/0076-6879(82)82089-2. [DOI] [PubMed] [Google Scholar]
- Taylor J. C., Crawford I. P. Purification and preliminary characterization of human leukocyte elastasel. Arch Biochem Biophys. 1975 Jul;169(1):91–101. doi: 10.1016/0003-9861(75)90320-3. [DOI] [PubMed] [Google Scholar]
- Twumasi D. Y., Liener I. E. Proteases from purulent sputum. Purification and properties of the elastase and chymotrypsin-like enzymes. J Biol Chem. 1977 Mar 25;252(6):1917–1926. [PubMed] [Google Scholar]
- Uhlenbruck G., Prokop O. An agglutinin from Helix pomatia, which reacts with terminal N-acetyl-D-galactosamine. Vox Sang. 1966 Jul-Aug;11(4):519–520. doi: 10.1111/j.1423-0410.1966.tb04250.x. [DOI] [PubMed] [Google Scholar]
- Widdicombe J. G. Control of secretion of tracheobronchial mucus. Br Med Bull. 1978 Jan;34(1):57–61. doi: 10.1093/oxfordjournals.bmb.a071460. [DOI] [PubMed] [Google Scholar]
- Wu R., Nolan E., Turner C. Expression of tracheal differentiated functions in serum-free hormone-supplemented medium. J Cell Physiol. 1985 Nov;125(2):167–181. doi: 10.1002/jcp.1041250202. [DOI] [PubMed] [Google Scholar]