Three molecular forms of a rat asialoglycoprotein receptor.
It has been reported that a rat asialoglycoprotein receptor is composed of three polypeptide chains with molecular masses of 43, 54, and 64 kilodaltons (43, 54, and 64-Kd forms) and that the first has a different primary structure from the latter two forms. Incorporation of [3H]leucine into these forms showed that no precursor-product relationship is found between the 54-Kd and 64-Kd forms. The half-life of the 43-Kd form (25 h) was shorter than those of the 54-Kd and 64-Kd forms (66 and 70 h, respectively). Glycopeptides of the three forms were prepared from rat livers previously labeled in vivo with [3H]glucosamine. Gel filtration analysis of the glycopeptides before and after endo H treatment revealed that they were all resistant to endo H. Alkali treatment did not change the elution position appreciately. These results indicate that the three molecular forms contained only complex oligosaccharide chains. The receptor was prepared from rat livers previously treated with tunicamycin in vivo and subjected to SDS-PAGE. A distinct band with a molecular mass of 33 Kd was observed. The receptor was also immunoprecipitated from rat hepatocytes in primary culture previously labeled with [35S]methionine and analysed by SDS-PAGE and fluorography. In addition to the major 43-Kd form, a band with a molecular mass of 41 Kd was found and tunicamycin treatment gave rise to a 33-Kd component, which is in good agreement with the receptor purified from tunicamycin treated rats. It is suggested that the 43-Kd form is synthesized as a 33 Kd polypeptide, cotranslationally glycosylated to form the 41 Kd component and then processed to the final 43-Kd form. We also think that the 43-Kd form could bind to asialoorosomucoid-Sepharose 4B without its carbohydrate chains.
H
H
Nakada
T
T
Sawamura
T
T
Okumura
Y
Y
Tashiro
D000818:Animals; D037263:Asialoglycoprotein Receptor; D004591:Electrophoresis, Polyacrylamide Gel; D005944:Glucosamine / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D008297:Male; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D011971:Receptors, Immunologic / Q000032:analysis* / Q000378:metabolism; D013462:Sulfur Radioisotopes; D014316:Tritium; D014415:Tunicamycin / Q000494:pharmacology
D037263:Asialoglycoprotein Receptor; D009844:Oligosaccharides; D011971:Receptors, Immunologic; D013462:Sulfur Radioisotopes; D014316:Tritium; D014415:Tunicamycin; D008715:Methionine; D007930:Leucine; D005944:Glucosamine