The LDL receptor in familial hypercholesterolemia: Use of human mutations to dissect a membrane protein

Since the discovery of the LDL receptor 13 years ago, a multidisciplinary approach to its study has revealed much about this important cell-surface protein. Most recently, we have developed tools in the form of full-length cDNAs and cloned genomic DNAs necessary to understand the molecular genetics of this locus. The frequent occurrence of mutations in the LDL receptor gene in patients with FH provides a fertile ground on which to explore the parts of the receptor that are necessary for its function. The analysis of four large deletions has revealed an unexpectedly universal involvement of Alu repeats in their generation. These studies indicate that repetitive DNAs can destabilize a gene through homologous recombination. Inasmuch as the LDL receptor gene is a mosaic of exons shared with at least five other proteins, it is possible that early exon-shuffling events involved recombination between these repetitive elements. Is it possible that the very plasticity that permitted evolution of the LDL receptor also accounts for its frequent disruption by mutation? Further study may help to answer this question. Mutations that disrupt the structure of the protein have been identified. The biochemical and cellular consequences of these mutations reveal crucial aspects of receptor structure. The receptor is clearly divided into quasi-independent domains with discrete functions. Mutations that disrupt the cytoplasmic domain alter the ability of the LDL receptor to cluster in coated pits, but they do not disrupt ligand binding or produce major effects on intracellular transport. Some of the mutations in the external domain disrupt binding but do not affect transport or internalization. Other mutations disrupt transport to the surface without abolishing binding. Some domains of the protein appear to function in a cooperative manner. Thus, deletions in the EGF precursor homology domain affect the ability of the receptor to bind LDL, indicating that this domain acts in collaboration with the actual amino-terminal binding domain. Clearly, further study of these human mutations will tell us much about the structure and function of the LDL receptor and probably other membrane proteins as well.