Mutational analysis of the ligand binding domain of the low density lipoprotein receptor

V. Esser, L. E. Limbird, M. S. Brown, J. L. Goldstein, D. W. Russell

Research output: Contribution to journalArticle

215 Citations (Scopus)

Abstract

The ligand binding domain of the low density lipoprotein (LDL) receptor contains seven imperfect repeats of a 40-amino acid cysteine-rich sequence. Each repeat contains clustered negative charges that have been postulated as ligand-binding sites. The adjacent region of the protein, the growth factor homology region, contains three cysteine-rich repeats (A-C) whose sequence differs from those in the ligand binding domain. To dissect the contribution of these different cysteine-rich repeats to ligand binding, we used oligonucleotide-directed mutagenesis to alter expressible cDNAs for the human LDL receptor which were then introduced into monkey COS cells by transfection. We measured the ability of the mutant receptors to bind LDL, which contains a single protein ligand for the receptor (apoB-100), and β-migrating very low density lipoprotein (β-VLDL), which contains apoB-100 plus multiple copies of another ligand (apoE). The results show that repeat 1 is not required for binding of either ligand. Repeats 2 plus 3 and repeats 6 plus 7 are required for maximal binding of LDL, but not β-VLDL. Repeat 5 is required for binding of both ligands. Repeat A in the growth factor homology region is required for binding of LDL, but not β-VLDL. These results support a model for the LDL receptor in which various repeats play additive roles in ligand binding, each repeat making a separate contribution to the binding event.

Original languageEnglish (US)
Pages (from-to)13282-13290
Number of pages9
JournalJournal of Biological Chemistry
Volume263
Issue number26
StatePublished - 1988

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LDL Receptors
Ligands
LDL Lipoproteins
Apolipoprotein B-100
Cysteine
Intercellular Signaling Peptides and Proteins
Mutagenesis
VLDL Lipoproteins
COS Cells
Apolipoproteins E
Site-Directed Mutagenesis
Oligonucleotides
Haplorhini
Transfection
Proteins
Complementary DNA
Binding Sites
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

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Mutational analysis of the ligand binding domain of the low density lipoprotein receptor. / Esser, V.; Limbird, L. E.; Brown, M. S.; Goldstein, J. L.; Russell, D. W.

In: Journal of Biological Chemistry, Vol. 263, No. 26, 1988, p. 13282-13290.

Research output: Contribution to journalArticle

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