Mucin biosynthesis and secretion in tracheal epithelial cells in primary culture
(2001) In Biochemical Journal 353(1). p.23-32- Abstract
Density-gradient centrifugation of bovine tracheal epithelial cell extracts revealed a 'high-density' (1.48 g/ml) sialic-acid-rich population as well as a 'low-density' (1.42 g/ml) one that reacted more strongly with a periodate-Schiff (PAS) assay. The sialic-acid-rich mucins were oligomeric molecules containing disulphide-bond-linked subunits and large glycosylated domains, whereas the PAS-reactive component seemed to be smaller and 'monomeric'. Only the 'high-density' population was secreted from cells cultured for 5 days on plastic or a collagen type 1, Matrigel or Vitrogen substrate. Release was less from cells grown on plastic than from those on a substrate and the amount was unaffected by increasing the thickness of the collagen... (More)
Density-gradient centrifugation of bovine tracheal epithelial cell extracts revealed a 'high-density' (1.48 g/ml) sialic-acid-rich population as well as a 'low-density' (1.42 g/ml) one that reacted more strongly with a periodate-Schiff (PAS) assay. The sialic-acid-rich mucins were oligomeric molecules containing disulphide-bond-linked subunits and large glycosylated domains, whereas the PAS-reactive component seemed to be smaller and 'monomeric'. Only the 'high-density' population was secreted from cells cultured for 5 days on plastic or a collagen type 1, Matrigel or Vitrogen substrate. Release was less from cells grown on plastic than from those on a substrate and the amount was unaffected by increasing the thickness of the collagen layer. For cells grown on collagen, the amount of the sialic-acid-rich mucin increased over 10 days, whereas the PAS-reactive component was largely absent after 24 h, which was consistent with an initial release of stored PAS-reactive molecules and synthesis of the sialic-acid-rich mucins de novo. Both [3H]proline and [35S]sulphate were poorly incorporated into mucins detected with the chemical assays but molecules with a higher buoyant density than that of either of the previously identified species were labelled with [35S]sulphate. The [35S]sulphate-labelled material yielded large trypsin-resistant fragments and contained O-linked glycans but was not affected by digestion with chondroitin ABC lyase or heparan sulphate lyase, suggesting that it is a mucin rather than a proteoglycan. [35S]Sulphate is thus a poor marker for the major oligomeric mucins produced by bovine tracheal epithelial cells but the radiolabel is incorporated into a heavily labelled mucin-like component.
(Less)
- author
- Svitacheva, N. LU and Davies, J. R. LU
- organization
- publishing date
- 2001
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Airway, Bovine trachea, Cell culture, Mucus, Radio-labelling
- in
- Biochemical Journal
- volume
- 353
- issue
- 1
- pages
- 23 - 32
- publisher
- Portland Press
- external identifiers
-
- scopus:0035152425
- pmid:11115395
- ISSN
- 0264-6021
- DOI
- 10.1042/bj3530023
- language
- English
- LU publication?
- yes
- id
- 4559df5f-8701-4830-bdca-17c346ecb511
- date added to LUP
- 2020-02-25 16:29:52
- date last changed
- 2024-01-02 06:16:18
@article{4559df5f-8701-4830-bdca-17c346ecb511,
abstract = {{<p>Density-gradient centrifugation of bovine tracheal epithelial cell extracts revealed a 'high-density' (1.48 g/ml) sialic-acid-rich population as well as a 'low-density' (1.42 g/ml) one that reacted more strongly with a periodate-Schiff (PAS) assay. The sialic-acid-rich mucins were oligomeric molecules containing disulphide-bond-linked subunits and large glycosylated domains, whereas the PAS-reactive component seemed to be smaller and 'monomeric'. Only the 'high-density' population was secreted from cells cultured for 5 days on plastic or a collagen type 1, Matrigel or Vitrogen substrate. Release was less from cells grown on plastic than from those on a substrate and the amount was unaffected by increasing the thickness of the collagen layer. For cells grown on collagen, the amount of the sialic-acid-rich mucin increased over 10 days, whereas the PAS-reactive component was largely absent after 24 h, which was consistent with an initial release of stored PAS-reactive molecules and synthesis of the sialic-acid-rich mucins de novo. Both [<sup>3</sup>H]proline and [<sup>35</sup>S]sulphate were poorly incorporated into mucins detected with the chemical assays but molecules with a higher buoyant density than that of either of the previously identified species were labelled with [<sup>35</sup>S]sulphate. The [<sup>35</sup>S]sulphate-labelled material yielded large trypsin-resistant fragments and contained O-linked glycans but was not affected by digestion with chondroitin ABC lyase or heparan sulphate lyase, suggesting that it is a mucin rather than a proteoglycan. [<sup>35</sup>S]Sulphate is thus a poor marker for the major oligomeric mucins produced by bovine tracheal epithelial cells but the radiolabel is incorporated into a heavily labelled mucin-like component.</p>}},
author = {{Svitacheva, N. and Davies, J. R.}},
issn = {{0264-6021}},
keywords = {{Airway; Bovine trachea; Cell culture; Mucus; Radio-labelling}},
language = {{eng}},
number = {{1}},
pages = {{23--32}},
publisher = {{Portland Press}},
series = {{Biochemical Journal}},
title = {{Mucin biosynthesis and secretion in tracheal epithelial cells in primary culture}},
url = {{http://dx.doi.org/10.1042/bj3530023}},
doi = {{10.1042/bj3530023}},
volume = {{353}},
year = {{2001}},
}