The University of Southampton
University of Southampton Institutional Repository

Size heterogeneity, phosphorylation and transmembrane organisation of desmosomal glycoproteins 2 and 3 (desmocollins) in MDCK cells

Size heterogeneity, phosphorylation and transmembrane organisation of desmosomal glycoproteins 2 and 3 (desmocollins) in MDCK cells
Size heterogeneity, phosphorylation and transmembrane organisation of desmosomal glycoproteins 2 and 3 (desmocollins) in MDCK cells
Metabolic labelling with [35S]methionine and immunoprecipitation with specific antibodies to bovine desmosomal glycoproteins 2 and 3 (dg2 and dg3: desmocollins) reveals a triplet of polypeptides of Mr 115,000, 107,000 and 104,000 in MDCK cells. Tunicamycin treatment shows that this heterogeneity does not arise through differential N-linked glycosylation. Under conditions in which cells are actively forming desmosomes, the largest polypeptide, dg2, becomes phosphorylated on serine, but the two smaller polypeptides, dg3a and 3b, do not. Controlled trypsinisation of intact cells yields three membrane-protected fragments (Mr 28,000, 24,000 and 23,000) derived from these glycoproteins. The largest of these fragments is phosphorylated but the two smaller fragments are not. A monoclonal antibody to bovine dg2 and dg3 stains MDCK cells cytoplasmically. In immunoblotting of MDCK cells the monoclonal antibody recognises dg2 strongly and shows a weaker reaction with a band of lower Mr corresponding to dg3a. It also recognises the immunoprecipitated 28,000 Mr fragment from trypsinised cells and a smaller fragment of 24,000 Mr. The simplest interpretation of these data is that all three glycoproteins have a transmembrane configuration with a single membrane-spanning domain, and show heterogeneity of size and phosphorylation in their cytoplasmic domains. The data are discussed in relation to the known structures of some cell adhesion molecules. Questions about the relative roles and distributions of the different polypeptides in desmosomal organisation are raised.
0021-9533
239-248
Parish, E.P.
cdc262b2-c32c-444e-95eb-1448165ef7dc
Collins, J.E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Mattey, D.L.
dff33b85-af6d-41c1-8fd5-582119fc15dd
Measures, H.R.
c41063df-9ddb-4e56-8549-aa90dc52614b
Venning, R.
d027c38f-e867-4c2c-82d5-3e80a5d3270d
Garrod, D.R.
b20b86ce-d168-4810-989f-7a3e11d122e2
Parish, E.P.
cdc262b2-c32c-444e-95eb-1448165ef7dc
Collins, J.E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Mattey, D.L.
dff33b85-af6d-41c1-8fd5-582119fc15dd
Measures, H.R.
c41063df-9ddb-4e56-8549-aa90dc52614b
Venning, R.
d027c38f-e867-4c2c-82d5-3e80a5d3270d
Garrod, D.R.
b20b86ce-d168-4810-989f-7a3e11d122e2

Parish, E.P., Collins, J.E., Mattey, D.L., Measures, H.R., Venning, R. and Garrod, D.R. (1990) Size heterogeneity, phosphorylation and transmembrane organisation of desmosomal glycoproteins 2 and 3 (desmocollins) in MDCK cells. Journal of Cell Science, 96 (2), 239-248. (PMID:2120245)

Record type: Article

Abstract

Metabolic labelling with [35S]methionine and immunoprecipitation with specific antibodies to bovine desmosomal glycoproteins 2 and 3 (dg2 and dg3: desmocollins) reveals a triplet of polypeptides of Mr 115,000, 107,000 and 104,000 in MDCK cells. Tunicamycin treatment shows that this heterogeneity does not arise through differential N-linked glycosylation. Under conditions in which cells are actively forming desmosomes, the largest polypeptide, dg2, becomes phosphorylated on serine, but the two smaller polypeptides, dg3a and 3b, do not. Controlled trypsinisation of intact cells yields three membrane-protected fragments (Mr 28,000, 24,000 and 23,000) derived from these glycoproteins. The largest of these fragments is phosphorylated but the two smaller fragments are not. A monoclonal antibody to bovine dg2 and dg3 stains MDCK cells cytoplasmically. In immunoblotting of MDCK cells the monoclonal antibody recognises dg2 strongly and shows a weaker reaction with a band of lower Mr corresponding to dg3a. It also recognises the immunoprecipitated 28,000 Mr fragment from trypsinised cells and a smaller fragment of 24,000 Mr. The simplest interpretation of these data is that all three glycoproteins have a transmembrane configuration with a single membrane-spanning domain, and show heterogeneity of size and phosphorylation in their cytoplasmic domains. The data are discussed in relation to the known structures of some cell adhesion molecules. Questions about the relative roles and distributions of the different polypeptides in desmosomal organisation are raised.

Full text not available from this repository.

More information

Published date: 1990

Identifiers

Local EPrints ID: 179791
URI: https://eprints.soton.ac.uk/id/eprint/179791
ISSN: 0021-9533
PURE UUID: 88d6e748-e0cd-4072-8306-01bda7f0cc03

Catalogue record

Date deposited: 04 Apr 2011 12:52
Last modified: 18 Jul 2017 12:02

Export record

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×