Domain map of the LDL receptor: Sequence homology with the epidermal growth factor precursor

Russell, David W.; Schneider, Wolfgang J.; Yamamoto, Tokuo; Luskey, Kenneth L.; Brown, Michael S.; Goldstein, Joseph L.

doi:10.1016/0092-8674(84)90388-x

Cited by 362 publications

(139 citation statements)

References 34 publications

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“…It should also be noted in this regard that the structure of the 8AR is quite different from that of several other membrane receptors whose structures are currently known. These include the receptors for low density lipoprotein (22), transferrin (23), Table 1 insulin (24), epidermal growth factor (25), and PDGF (26). In each case these receptors possess only a single membranespanning domain.…”

Section: Resultsmentioning

confidence: 99%

cDNA for the human beta 2-adrenergic receptor: a protein with multiple membrane-spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth factor.

Kobilka¹,

Dixon²,

Frielle³

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

606

184

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We have isolated and sequenced a cDNA encoding the human 132-adrenergic receptor. The deduced amino acid sequence (413 residues) is that of a protein containing seven clusters of hydrophobic amino acids suggestive of membrane-spanning domains. While the protein is 87% identical overall with the previously cloned hamster P2-adrenergic receptor, the most highly conserved regions are the putative transmembrane helices (95% identical) and cytoplasmic loops (93% identical), suggesting that these regions of the molecule harbor important functional domains. Several of the transmembrane helices also share lesser degrees of identity with comparable regions of select members of the opsin family of visual pigments. We have localized the gene for the P2-adrenergic receptor to q31-q32 on chromosome 5. This is the same position recently determined for the gene encoding the receptor for platelet-derived growth factor and is adjacent to that for the FMS protooncogene, which encodes the receptor for the macrophage colony-stimulating factor.Many hormones, neurotransmitters, and drugs influence cellular metabolic activities by stimulating the adenylate cyclase system, leading to the generation of the second messenger cAMP and activation of the cAMP-dependent protein kinase. The molecular components of this plasma membrane signaling system include specific receptors that bind ligands, the catalyst that converts ATP to cAMP, and guanine nucleotide regulatory or G proteins that functionally couple the receptors to the enzyme (1). The latter two components of the system have been purified and genes encoding several members of the "G protein family" have been cloned.Of the receptors that are coupled to adenylate cyclase the only one that has been characterized in any detail is the p-adrenergic receptor (,8AR). Two pharmacologically and physiologically distinct subtypes of this receptor, termed ,31AR and f32AR, are both membrane glycoproteins of Mr 64,000 (2). Very recently, we reported cloning of cDNA and the gene for the hamster f32AR (3). The deduced protein sequence indicated a protein of 418 amino acids, with seven clusters of hydrophobic amino acids likely representing membrane-spanning regions. This topology resembles that of the visual "light receptor" rhodopsin, which also possesses seven membrane-spanning domains (4-6).We now report the cloning and complete nucleotide sequence of the cDNA for the human ,82AR. While the receptor is highly similar to its hamster counterpart (87% of the amino acid residues are identical), significant regional differences in the extent of identity are noted. METHODScDNA Library Screening. The human placenta cDNA library was kindly provided by Evan Sadler (Washington University School of Medicine). The cDNA was prepared from term placenta poly(A)+ RNA and cloned in phage Xgtll. The library contains 5 x 106 independent recombinants. The A431 Xgtll library was prepared from poly(A)+ RNA from actively dividing A431 cells by methods previously described (3). It contains 1 x 106 independent reco...

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Section: Resultsmentioning

confidence: 99%

cDNA for the human beta 2-adrenergic receptor: a protein with multiple membrane-spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth factor.

Kobilka¹,

Dixon²,

Frielle³

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

606

184

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“…IgG-C7, a mouse monoclonal antibody that recognizes an epitope in the first cysteine-rich repeat of the binding domain of the human LDL receptor (7), was prepared and radiolabeled with 1251 as described (2). A polyclonal anti-LDL receptor antibody, 698-7, was raised against purified bovine LDL receptor (11). B3/25, a mouse monoclonal antibody to the human transferrin receptor, was provided by I. S. Trowbridge (The Salk Institute for Biological Studies, San Diego, CA).…”

Section: Methodsmentioning

confidence: 99%

Deletion in the first cysteine-rich repeat of low density lipoprotein receptor impairs its transport but not lipoprotein binding in fibroblasts from a subject with familial hypercholesterolemia.

Leitersdorf

Hobbs

Fourie

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

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The ligand-binding domain of the low density lipoprotein (LDL) receptor is composed of seven cysteine-rich repeats, each -40 amino acids long. Previous studies by van Driel et al. [van Driel, I. R., Goldstein, J. L., Sudhof, T. C. & Brown, M. S. (1987) J. Biol. Chem. 262, 17443-17449] showed that if the first repeat of the ligand-binding domain (encoded by exon 2) is deleted, the receptor fails to bind an anti-LDL receptor monoclonal antibody (IgG-C7) but continues to bind LDL with high affinity. Cultured fibroblasts from a Black South African Xhosa patient (TT) with the clinical syndrome of homozygous familial hypercholesterolemia demonstrated highaffinity cell-surface binding of 125I-labeled LDL but not '25I-labeled IgG-C7. Previous haplotype analysis, using 10 restriction fragment length polymorphic sites, suggested that the patient inherited two identical LDL receptor alleles. The polymerase chain reaction technique was used to selectively amplify exon 2 of the LDL receptor gene from this patient. Sequence analysis of the amplified fragment disclosed a deletion of six base pairs that removes two amino acids, aspartic acid and glycine, from the first cysteine-rich ligand binding repeat. The mutation creates a new Pst I restriction site that can be used to detect the deletion. The existence of this mutant allele confirms that the epitope of IgG-C7 is located in the first cysteine-rich repeat and that this repeat is not necessary for LDL binding. The mutant gene produced a normally sized 120-kilodalton LDL receptor precursor protein that matured to the 160-kilodalton form at less than one-fourth the normal rate. Thus, deletion of two amino acids within the first cysteine-rich repeat retards receptor transport from the endoplasmic reticulum to the cell surface, in contrast to deletion of the entire first repeat, which has no effect on receptor maturation.Mutations in the low density lipoprotein (LDL) receptor gene result in the autosomal dominant disease familial hypercholesterolemia (FH) (1). The LDL receptor is a transmembrane protein that binds LDL and mediates its uptake into the cell. Individuals with two mutant LDL receptor alleles have profoundly fewer or no functional LDL receptors on their cell surfaces and this is associated with markedly elevated plasma LDL-cholesterol levels. These individuals develop premature coronary atherosclerosis that often leads to myocardial infarctions in early childhood. In homozygous FH, mutations in each LDL receptor allele can be either identical (a true homozygote) or different (a compound heterozygote).

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“…Starting from a cDNA mixture obtained by reverse transcription of brain microvessels mRNAs, the bovine LDL-R cDNA [29,30] was amplified by using BTLDLP19 (5'-TGCACTCCATCTCCAGCATC-G-3') as forward primer and BTLDLM3 (5'-GAGGCAGCTTCT-CATGTCCTTGG-3') as backward primer (Fig. 1A); an amplified fragment of 392 bp was obtained.…”

Section: Pcr Strategy and Restriction Analysismentioning

confidence: 99%

Expression of receptors for native and chemically modified low‐density lipoproteins in brain microvessels

et al. 1997

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Despite the importance of cholesterol metabolism in the central nervous system, only relatively few studies have dealt with the cerebral uptake and transport of lipids into the brain compartment. These functions are mediated by the endothelium of brain microvessels, which forms the anatomical basis of the blood-brain barrier. By a reverse transcriptase PCR study of messenger RNA expression we could show, in bovine brain microvessels, the presence of transcripts of native low-density lipoprotein receptor and of both type I and II scavenger receptors. Brain microvessels therefore appear to play an active role in the uptake of native and modified low-density lipoproteins.

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Domain map of the LDL receptor: Sequence homology with the epidermal growth factor precursor

Cited by 362 publications

References 34 publications

cDNA for the human beta 2-adrenergic receptor: a protein with multiple membrane-spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth factor.

cDNA for the human beta 2-adrenergic receptor: a protein with multiple membrane-spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth factor.

Deletion in the first cysteine-rich repeat of low density lipoprotein receptor impairs its transport but not lipoprotein binding in fibroblasts from a subject with familial hypercholesterolemia.

Expression of receptors for native and chemically modified low‐density lipoproteins in brain microvessels

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