Document Detail


Mucin biosynthesis and secretion in tracheal epithelial cells in primary culture.
MedLine Citation:
PMID:  11115395     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 [(3)H]proline and [(35)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 [(35)S]sulphate. The [(35)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. [(35)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.
Authors:
N Svitacheva; J R Davies
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  353     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  2001 Jan 
Date Detail:
Created Date:  2001-01-26     Completed Date:  2001-02-08     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  England    
Other Details:
Languages:  eng     Pagination:  23-32     Citation Subset:  IM    
Affiliation:
Department of Cell and Molecular Biology, Section for Molecular Pathogenesis, Lund University, P.O. Box 94, S-22100 Lund, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Alkylation
Animals
Cattle
Cell Extracts
Cells, Cultured
Centrifugation, Isopycnic
Centrifugation, Zonal
Epithelial Cells / cytology,  metabolism*,  secretion
Kinetics
Molecular Weight
Mucins / biosynthesis*,  chemistry,  metabolism,  secretion*
N-Acetylneuraminic Acid / metabolism
Reducing Agents / metabolism
Trachea / cytology,  metabolism*,  secretion
Trypsin / metabolism
Chemical
Reg. No./Substance:
0/Cell Extracts; 0/Mucins; 0/Reducing Agents; 131-48-6/N-Acetylneuraminic Acid; EC 3.4.21.4/Trypsin
Comments/Corrections

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