Synthesis and secretion of mucin by the human colonic tumour cell line LS180.

Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Research Abstract Details 

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Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Abstract Text:

D J McCool,J F Forstner,G G Forstner,

Pulse-chase labelling experiments were performed using the mucin-producing colonic carcinoma cell line LS180. Cells were pulsed with [3H]threonine or [3H]glucosamine and chased in complete media without radiolabel for various lengths of time. From cell and media extracts obtained at each time point, mucin proteins were immunoprecipitated with specific anti-mucin antibodies and analysed by SDS/PAGE and fluorography. At short labelling times with [3H]threonine, without chase, a monomeric thiol-reduction-resistant mucin precursor of apparent molecular mass > 670 kDa was identified. The precursor, in contrast to oligomeric species, was not labelled by [3H]glucosamine but exhibited binding to Vicia villosa isolectin B4, suggesting the presence of some core GalNAc residues. Treatment with tunicamycin to inhibit N-glycosylation had no effect on the apparent mass of the precursor. Identity of the mucin antigen with MUC2 mucin was established by immunoprecipitation with antibodies specific for a MUC2 tandem repeat and C-terminal regions. With increasing chase time the precursor was replaced by thiol-reduction-sensitive mucin oligomers that reached peak intracellular radiolabelling with [3H]threonine by 2 h of chase, and then declined. Only oligomeric mucin was secreted into the medium. Secretion of [3H]threonine-labelled mucin was detectable after 2 h of chase and increased as the cytoplasmic mucin label declined. Monensin inhibited [3H]glucosamine incorporation, sialylation and baseline (non-regulated) mucin secretion without affecting initial [3H]threonine incorporation or oligomerization. Oligomerization and Golgi transport are therefore essential early steps in MUC2 mucin secretion. Oligomerization may follow some core O-glycosylation with GalNAc, but precedes elongation of oligosaccharide chains.

Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Publishing Authors By Initials

DJ McCool,JF Forstner,GG Forstner,

For similar cells: cells, cultured: tumor cells, cultured research abstracts see: cells: cells, cultured: tumor cells, cultured research

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Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: The Biochemical journal

VOLUME: 302 ( Pt 1)

Page Numbers: 111-8

Journal Abbreviation: Biochem. J.

ISSN: 0264-6021

DAY: 15

MONTH: Aug

YEAR: 1994

Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 2984726

Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Keywords Mesh Terms:

KEYWORDS: Tumor Cells, Cultured

MESH TERMS: biosynthesis

Chemical & Substance for Abstract: Synthesis and secretion of mucin by the human colonic tumour cell line LS180. Information

Substance Name: Neuraminidase

Registry Number: EC 3.2.1.18

Grant and Affiliation Information for Synthesis and secretion of mucin by the human colonic tumour cell line LS180.

AFFILIATION: Hospital for Sick Children, Toronto, Ontario, Canada.

Country: ENGLAND

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MEDLINETA: Biochem J

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