Affinity labeling of the galactose/N-acetylgalactosamine-specific receptor of rat hepatocytes: preferential labeling of one of the subunits.
The galactose/N-acetylgalactosamine-specific receptor (also known as asialoglycoprotein receptor) of rat hepatocytes consists of three subunits, one of which [43 kilodalton (kDa)] exists in a greater abundance (up to 70% of total protein) over the two minor species (52 and 60 kDa). When the receptor on the hepatocyte membranes was photoaffinity labeled with an 125I-labeled high-affinity reagent [a triantennary glycopeptide containing an aryl azide group on galactosyl residues; Lee, R. T., & Lee, Y. C. (1986) Biochemistry 25, 6835-6841], the labeling occurred mainly (51-80%) on one of the minor bands (52 kDa). Similarly, affinity-bound, N-acetylgalactosamine-modified lactoperoxidase radioiodinated the same 52-kDa band preferentially. In contrast, both the photoaffinity labeling and lactoperoxidase-catalyzed iodination of the purified, detergent-solubilized receptor resulted in a distribution of the label that is comparable to the Coomassie blue staining pattern of the three bands; i.e., the 43-kDa band was the major band labeled. These and other experimental results suggest that the preferential labeling of the minor band and inefficient labeling of the major band on the hepatocyte membrane resulted from a specific topological arrangement of these subunits on the membranes. We postulate that in the native, membrane-bound state of the receptor, the 52-kDa minor band is topologically prominent, while the major (43 kDa) band is partially masked. This partial masking may result from a tight packing of the receptor subunits on the membranes to form a lattice work [Hardy, M. R., Townsend, R. R., Parkhurst, S. M., & Lee, Y. C. (1985) Biochemistry 24, 22-28].
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
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D000345:Affinity Labels / Q000378:metabolism*; D000818:Animals; D037263:Asialoglycoprotein Receptor; D002478:Cells, Cultured; D007457:Iodine Radioisotopes; D008099:Liver / Q000276:immunology*; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D010777:Photochemistry; D051381:Rats; D011919:Rats, Inbred Strains; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism*
D000345:Affinity Labels; D037263:Asialoglycoprotein Receptor; D007457:Iodine Radioisotopes; D046911:Macromolecular Substances; D011971:Receptors, Immunologic