Faculty Collaboration Database

Influence of quaternary structure on glycosylation. Differential subunit association affects the site-specific glycosylation of the common beta-chain from Mac-1 and LFA-1. J Biol Chem 1986 Oct 05;261(28):13186-96

Date

10/05/1986

Pubmed ID

3531197

Scopus ID

2-s2.0-0022898398 (requires institutional sign-in at Scopus site)   60 Citations

Abstract

The influence of quaternary structure on glycosylation was evaluated in a macrophage-like cell line, P388D1. This cell line simultaneously synthesizes two structurally related glycoproteins, Mac-1 and LFA-1. Mac-1 and LFA-1 each contain two subunits in noncovalent association in an alpha 1 beta 1 structure. The beta-chain polypeptides of these two glycoproteins have identical primary structures while their alpha-chain polypeptides are distinct. For both Mac-1 and LFA-1, the association of the alpha- and beta-chains occurs prior to any Golgi-mediated processing of the oligosaccharide moieties on either one of the subunits. To evaluate the effects of differential subunit association on the site-specific glycosylation of the beta-chain, [3H]glucosamine-labeled oligosaccharides were isolated from the beta-chain of Mac-1 and LFA-1 and were compared by a variety of enzymatic and chromatographic techniques. Reverse-phase high performance liquid chromatography analyses of tryptic-chymotryptic glycopeptides suggest that each beta-chain has at least five glycosylation sites. Structural analysis of oligosaccharides from each corresponding glycopeptide fraction of the beta-chains of Mac-1 or LFA-1 (comparing their glycosidase sensitivities, behavior on serial lectin affinity chromatography, size heterogeneity, extent of sialylation, and branching) indicates that the LFA-1 beta-chain is glycosylated substantially differently on at least four of its sites, compared to the corresponding sites of the Mac-1 beta-chain, even though they are simultaneously synthesized in the same cells. Thus, these data demonstrate that quaternary structure can influence the site-specific glycosylation of a protein, even when the polypeptide structure and the cellular glycosylation machinery remain constant.

Author List

Dahms NM, Hart GW

Author

Nancy M. Dahms PhD Professor in the Biochemistry department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Antigens, Surface
Binding Sites
Cell Line
Chromatography, Affinity
Chromatography, Gel
Chromatography, High Pressure Liquid
Electrophoresis, Polyacrylamide Gel
Glycosylation
Leukemia P388
Lymphocyte Function-Associated Antigen-1
Macromolecular Substances
Macrophage-1 Antigen
Mice
Oligosaccharides