title: The oligomerization domain of the asialoglycoprotein receptor preferentially forms 2:2 heterotetramers in vitro.
abstract: The human hepatic asialoglycoprotein receptor is a noncovalent hetero-oligomer composed of two homologous subunits, H1 and H2, with an as yet unknown stoichiometry. Ligand specificity and binding affinity depend on the arrangement of the subunits in the complex. An 80-amino acid segment connecting the transmembrane and the carbohydrate binding domains contains heptad repeats characteristic of alpha-helical coiled coil structure. We expressed and purified corresponding peptides, H1S and H2S, and confirmed by circular dichroism spectroscopy that they can assume alpha-helical conformation. Oxidative cross-linking of amino-terminal cysteines generated specific covalent oligomers, indicating that separately H1S forms trimers and H2S tetramers. Upon mixing, covalent heterotetramers were formed with a preferred stoichiometry of 2 H1S and 2 H2S peptides. These results suggest that the stalk segments of the receptor subunits oligomerize to constitute an alpha-helical coiled coil stalk on top of which the carbohydrate binding domains are exposed for ligand binding. We propose that the functional asialoglycoprotein receptor is a 2:2 heterotetramer.
authors.affiliation: Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland. spiess@ubaclu.unibas.ch
authors.forename: M D
authors.initials: MD
authors.lastname: Bider
authors.forename: J M
authors.initials: JM
authors.lastname: Wahlberg
authors.forename: R A
authors.initials: RA
authors.lastname: Kammerer
authors.forename: M
authors.initials: M
authors.lastname: Spiess
mesh_terms: D000595:Amino Acid Sequence; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins / Q000378:metabolism*; D002942:Circular Dichroism; D005455:Fluorescent Antibody Technique; D006801:Humans; D008099:Liver / Q000737:chemistry; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism
chemical_list: D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D011956:Receptors, Cell Surface