The University of Southampton
University of Southampton Institutional Repository

Site-specific glycosylation of recombinant rat and human soluble CD4 variants expressed in Chinese hamster ovary cells

Site-specific glycosylation of recombinant rat and human soluble CD4 variants expressed in Chinese hamster ovary cells
Site-specific glycosylation of recombinant rat and human soluble CD4 variants expressed in Chinese hamster ovary cells

The rat and human forms of the T-cell surface glycoprotein CD4 share a common glycosylation site at the Asn270/271 position but differ with respect to the locations of the second glycosylation sites at Asn159 (rat) and Asn300 (human). The glycosylation of soluble recombinant forms of human and rat CD4 (sCD4) expressed in Chinese hamster ovary cells has been characterized. The most obvious differences between the rat and human sCD4 oligosaccharides were the greater abundance of oligomannose and hybrid oligosaccharides on rat sCD4 and the presence of oligosaccharides carrying a terminal alpha-galactose residue on human sCD4. This is the first report of the occurrence of alpha-galactose residues on a glycoprotein expressed in Chinese hamster ovary cells. Comparison of mutant rat sCD4 molecules with single glycosylation sites and glycopeptides indicated that site-specific and independent processing occurred at each glycosylation site. The glycosylation at the conserved site at Asn270 of rat sCD4 was identical to that seen for the equivalent site in human sCD4, and the oligomannose and hybrid structures were restricted to the nonconserved site at Asn159 in rat sCD4. However, there was more oligosaccharide processing at this site in a truncated form of rat sCD4 consisting of the two NH2-terminal domains. These results indicate that not only the local three-dimensional structure but also domain interactions can influence the processing at individual glycosylation sites.

Amino Acid Sequence, Animals, CD4 Antigens, CHO Cells, Carbohydrate Conformation, Carbohydrate Sequence, Chromatography, Gel, Cricetinae, Glycopeptides, Glycosylation, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Oligosaccharides, Protein Processing, Post-Translational, Rats, Recombinant Proteins, Transfection, Journal Article, Research Support, Non-U.S. Gov't
0021-9258
3260-3267
Ashford, D.A.
96b706e4-b6a0-4ec0-bf41-03632378b24a
Alafi, Christopher D.
993c5fdd-9132-4cae-adba-4ae612472fd7
Gamble, Victoria M.
023fbe1c-28f0-40e8-85b5-36be10abfecf
Mackay, D.J.
588a653e-9785-4a00-be71-4e547850ee4a
Rademacher, Thomas W.
9bf5b14b-b6ab-4c1a-8b58-6183466ec41a
Williams, Phillip J.
ee6edd56-cd74-411f-886a-61b500d8f12c
Dwek, Raymond A.
d8d9d5f8-f2c0-414e-b6b0-df77a33f0da4
Barclay, A. Neil
5199192a-78a1-4a00-946c-bced641ee10a
Davis, Simon J.
77c9e91c-a2c6-498a-9128-bf164a8da8de
Somoza, Chamorro
32027c4d-e0a1-43a5-88d2-03563b48b322
Ward, Harry A.
88c4596e-dcab-47bb-bccb-fc361fc46adf
Williams, Alan F.
54c2f49c-dfe6-4922-8c92-8fdd9a303684
Ashford, D.A.
96b706e4-b6a0-4ec0-bf41-03632378b24a
Alafi, Christopher D.
993c5fdd-9132-4cae-adba-4ae612472fd7
Gamble, Victoria M.
023fbe1c-28f0-40e8-85b5-36be10abfecf
Mackay, D.J.
588a653e-9785-4a00-be71-4e547850ee4a
Rademacher, Thomas W.
9bf5b14b-b6ab-4c1a-8b58-6183466ec41a
Williams, Phillip J.
ee6edd56-cd74-411f-886a-61b500d8f12c
Dwek, Raymond A.
d8d9d5f8-f2c0-414e-b6b0-df77a33f0da4
Barclay, A. Neil
5199192a-78a1-4a00-946c-bced641ee10a
Davis, Simon J.
77c9e91c-a2c6-498a-9128-bf164a8da8de
Somoza, Chamorro
32027c4d-e0a1-43a5-88d2-03563b48b322
Ward, Harry A.
88c4596e-dcab-47bb-bccb-fc361fc46adf
Williams, Alan F.
54c2f49c-dfe6-4922-8c92-8fdd9a303684

Ashford, D.A., Alafi, Christopher D., Gamble, Victoria M., Mackay, D.J., Rademacher, Thomas W., Williams, Phillip J., Dwek, Raymond A., Barclay, A. Neil, Davis, Simon J., Somoza, Chamorro, Ward, Harry A. and Williams, Alan F. (1993) Site-specific glycosylation of recombinant rat and human soluble CD4 variants expressed in Chinese hamster ovary cells. The Journal of Biological Chemistry, 268 (5), 3260-3267.

Record type: Article

Abstract

The rat and human forms of the T-cell surface glycoprotein CD4 share a common glycosylation site at the Asn270/271 position but differ with respect to the locations of the second glycosylation sites at Asn159 (rat) and Asn300 (human). The glycosylation of soluble recombinant forms of human and rat CD4 (sCD4) expressed in Chinese hamster ovary cells has been characterized. The most obvious differences between the rat and human sCD4 oligosaccharides were the greater abundance of oligomannose and hybrid oligosaccharides on rat sCD4 and the presence of oligosaccharides carrying a terminal alpha-galactose residue on human sCD4. This is the first report of the occurrence of alpha-galactose residues on a glycoprotein expressed in Chinese hamster ovary cells. Comparison of mutant rat sCD4 molecules with single glycosylation sites and glycopeptides indicated that site-specific and independent processing occurred at each glycosylation site. The glycosylation at the conserved site at Asn270 of rat sCD4 was identical to that seen for the equivalent site in human sCD4, and the oligomannose and hybrid structures were restricted to the nonconserved site at Asn159 in rat sCD4. However, there was more oligosaccharide processing at this site in a truncated form of rat sCD4 consisting of the two NH2-terminal domains. These results indicate that not only the local three-dimensional structure but also domain interactions can influence the processing at individual glycosylation sites.

Full text not available from this repository.

More information

Published date: 15 February 1993
Keywords: Amino Acid Sequence, Animals, CD4 Antigens, CHO Cells, Carbohydrate Conformation, Carbohydrate Sequence, Chromatography, Gel, Cricetinae, Glycopeptides, Glycosylation, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Oligosaccharides, Protein Processing, Post-Translational, Rats, Recombinant Proteins, Transfection, Journal Article, Research Support, Non-U.S. Gov't

Identifiers

Local EPrints ID: 418607
URI: https://eprints.soton.ac.uk/id/eprint/418607
ISSN: 0021-9258
PURE UUID: 741d7a74-6101-46a8-9f12-ad79b1a77837
ORCID for D.J. Mackay: ORCID iD orcid.org/0000-0003-3088-4401

Catalogue record

Date deposited: 12 Mar 2018 17:30
Last modified: 14 Mar 2019 01:48

Export record

Contributors

Author: D.A. Ashford
Author: Christopher D. Alafi
Author: Victoria M. Gamble
Author: D.J. Mackay ORCID iD
Author: Thomas W. Rademacher
Author: Phillip J. Williams
Author: Raymond A. Dwek
Author: A. Neil Barclay
Author: Simon J. Davis
Author: Chamorro Somoza
Author: Harry A. Ward
Author: Alan F. Williams

University divisions

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.

×