Abnormal structure and co-operative binding of low-density lipoprotein receptors containing the Glu-80-->Lys mutation

The properties of low-density lipoprotein (LDL) receptors containing a Glu to Lys substitution at position 80 have been studied in fibroblasts from a homozygous familial hypercholesterolaemic subject (MB) and in monkey COS cells transfected with the mutant cDNA. Receptors containing the Glu-80-->Lys mutation were processed more slowly than the normal protein and only approx. 50% reached the surface as the mature form. Both cell types exhibited a normal concentration binding curve for beta-very-low-density lipoproteins (beta-VLDL) but an atypical, sigmoidal curve for LDL. The mature mutant receptor protein migrated abnormally slowly on SDS-PAGE under non-reducing conditions but normally under reducing conditions or after treatment with neuraminidase. It also showed an unusual ability to form dimers that were stable in detergents. Transfected normal and mutant receptors were apparently cleaved on the surface of the cells to give a product lacking the NH2-terminal portion of the protein, which was resistant to further proteolytic digestion. The results suggest that the Glu-80-->Lys substitution produces a change in the conformation of the protein, stabilized by polysaccharide chains, which results in a strong self-association of receptor molecules that affects their ability to bind LDL.