Three types of low density lipoprotein receptor-deficient mutant have pleiotropic defects in the synthesis of N-linked, O-linked, and lipid-linked carbohydrate chains.
Biochemical, immunological, and genetic techniques were used to investigate the genetic defects in three types of low density lipoprotein (LDL) receptor-deficient hamster cells. The previously isolated ldlB, ldlC, and ldlD mutants all synthesized essentially normal amounts of a 125,000-D precursor form of the LDL receptor, but were unable to process this receptor to the mature form of 155,000 D. Instead, these mutants produced abnormally small, heterogeneous receptors that reached the cell surface but were rapidly degraded thereafter. The abnormal sizes of the LDL receptors in these cells were due to defective processing of the LDL receptor's N- and O-linked carbohydrate chains. Processing defects in these cells appeared to be general since the ldlB, ldlC, and ldlD mutants also showed defective glycosylation of a viral glycoprotein, alterations in glycolipid synthesis, and changes in resistance to several toxic lectins. Preliminary structural studies suggested that these cells had defects in multiple stages of the Golgi-associated processing reactions responsible for synthesis of glycolipids and in the N-linked and O-linked carbohydrate chains of glycoproteins. Comparisons between the ldl mutants and a large number of previously isolated CHO glycosylation defective mutants showed that the genetic defects in ldlB, ldlC, and ldlD cells were unique and that only very specific types of carbohydrate alteration could dramatically affect LDL receptor function.
D M
DM
Kingsley
K F
KF
Kozarsky
M
M
Segal
M
M
Krieger
D000818:Animals; D002240:Carbohydrate Sequence; D002451:Cell Compartmentation; D006224:Cricetinae; D006017:Glycolipids / Q000378:metabolism*; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D008970:Molecular Weight; D009154:Mutation; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D011973:Receptors, LDL / Q000235:genetics* / Q000276:immunology; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000378:metabolism
C028391:G protein, vesicular stomatitis virus; D006017:Glycolipids; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011973:Receptors, LDL; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D014415:Tunicamycin