A Receptor-Mediated Pathway for Cholesterol Homeostasis

Brown, Michael S.; Goldstein, Joseph L.

doi:10.1126/science.3513311

Cited by 5,174 publications

(2,826 citation statements)

References 146 publications

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“…It has also been observed that statins up-regulate hepatic LDLR expression and inhibition of HMG-CoA reductase results in a secondary increase in LDLR activity (3,24,25). Therefore, it is possible that statin therapy might affect the results even though peripheral lymphocytes, and not hepatocytes, were used in this study (26,27).…”

Section: Discussionmentioning

confidence: 96%

A novel method for determining functional LDL receptor activity in familial hypercholesterolemia: Application of the CD3/CD28 assay in lymphocytes

Tada

Kawashiri

Noguchi

et al. 2009

Clinica Chimica Acta

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Abbreviated title: An improved assay for LDL receptor activity using anti-CD3/CD28 beads Abbreviations: DiI, 3,3"--dioctadecylindocarbocyanin; FH, familial hypercholesterolemia; NARC-1, neural apoptosis-regulated convertase 1; FITC, fluorescein isothiocyanate; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; LDLR, low-density lipoprotein receptor; LPDS, lipoprotein deficient serum; rIL-2, recombinant interleukin-2; MF, mean fluorescence; TC, total cholesterol; CV, coefficient of variation. 3 AbstractBackground: The objective of this study was to develop a new and simple method for measuring low-density lipoprotein receptor (LDLR) activity using peripheral lymphocytes enabling us to clinically diagnose familial hypercholesterolemia (FH) and ascertain the involved mutations (such as K790X mutation), that might not be clearly detected in the conventional method.Methods: Our method comprised the following 2 features: First, we used anti-CD3/CD28 beads to stimulate T-lymphocytes to obtain a uniform fraction of lymphocytes and maximum up-regulation of LDLR. Second, we excluded the possibility of overestimation of lymphocyte signals bound only to its surface, by adding heparin to the cultured lymphocytes used for the LDLR assay.Results: Based on the genetic mutation, the FH subjects were divided into two groups, K790X, (n = 20) and P664L, (n = 5), and their LDLR activities was measured by this method, which was found to be 55.3 ± 8.9% and 63.9 ± 13.8%, respectively, of that of the control group (n = 15). In comparison, the LDLR activity was 86.1 ± 11.6% (K790X) and 73.3 ± 6.3% (P664L) of that of the control group when measured by the conventional method, indicating that impairment of LDLR function in FH K790X subjects was much more clearly differenciated with our method than with the conventional method (paired t-test, p < 0.0001). The levels of LDLR expression also showed similar tendencies, that is, 89.4 ± 13.2% (K790X) and 76.9 ± 17.4% (P664L) of that of the control group when measured by the conventional method, and 78.1 ± 9.7% (K790X) and 70.3 ± 26.5% (P664L) when measured by our new method. In addition, we confirmed that there was little influence of statin treatment on LDLR activity among the study subjects when our method was used.Conclusion: These results demonstrate that our new method is applicable for measuring LDLR 4 activity, even in subjects with an internally defective allele, and that T-lymphocytes of the FH K790X mutation possess characteristics of that allele.

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Section: Discussionmentioning

confidence: 96%

A novel method for determining functional LDL receptor activity in familial hypercholesterolemia: Application of the CD3/CD28 assay in lymphocytes

Tada

Kawashiri

Noguchi

et al. 2009

Clinica Chimica Acta

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“…Therefore mutations in the APOB 100 will drastically alter its functional activity leading to a decrease in its binding to LDLR, thereby delaying the clearance of LDL particles. The latter situation usually results in a mild or severe form of hypercholesterolemia together with an increased risk for early onset atherosclerosis [10,11]. In contrast to LDLR, only a small number of functional mutations have been identified in APOB gene such as R3500Q [12], R3500 W [13], and R3531C [14].…”

Section: Introductionmentioning

confidence: 99%

Molecular Characterization of Iranian Patients with Possible Familial Hypercholesterolemia

et al. 2011

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Familial hypercholesterolemia (FH) is an autosomal dominant disorder of lipoprotein metabolism caused mainly by mutations in the low-density lipoprotein receptor (LDLR) and apolipoprotein B 100 (APOB) genes. Until now, the molecular basis of FH has been demonstrated in detail in many populations, but there is still very limited Molecular data concerning FH in Iran. The aim of this study was to characterize the LDLR and APOB gene mutations in an Iranian population. A total of 30 non-related Iranian possible FH subjects were studied. Diagnosis of FH was based on the Dutch Lipid Clinic Network diagnostic criteria. All samples were initially tested for three common APOB gene mutations including R3500Q, R3500 W and R3531C using PCR-RFLP assay. Subsequently, promoter and coding region of the LDLR gene was screened by PCR-SSCP analysis and positive results were confirmed by DNA sequencing. Four previously reported polymorphisms 1413G [ A, 1725C [ T, 1773T [ C and 2140 ? 5G [ A were found in *17% (5/30) of population studied.Moreover, no variation was found in APOB gene. Our data indicated that LDLR and APOB gene mutations have not contribution to possible FH in Iranian population studied here. However, we examined three common APOB mutations and LDLR in only 30 patients, and to determine the role of these genes in developing FH in Iran, more FH samples and populations needed to be investigated for the mutations of the related genes.

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“…Classical studies by Brown and Goldstein revealed that the low-density lipoprotein receptor (LDLR) pathway plays a key role in plasma cholesterol homeostasis [1]. Following discovery of the LDLR, a growing number of homologous receptors have been identified [2].…”

Section: Introductionmentioning

confidence: 99%

Apolipoprotein E isoform-specific binding to the low-density lipoprotein receptor

Yamamoto¹,

Choi²,

Ryan³

2008

Analytical Biochemistry

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Apolipoprotein E (apoE) is a ligand for members of the low-density lipoprotein receptor (LDLR) family and functions in plasma cholesterol homeostasis. A fluorescence-based assay has been employed in molecular studies of receptor-ligand interactions. Competition experiments revealed isoform specific differences in binding of lipid-associated apoE N terminal (NT) domain to a recombinant soluble LDLR (sLDLR). In a similar manner, lipid associated, but not lipid-free, fulllength apoE3 showed binding activity to sLDLR. The molecular chaperone, Receptor Associated Protein, inhibited apoE3-NT-phospholipid complex binding to sLDLR. Kinetic studies of apoE3-NT-phospholipid complex interaction with sLDLR revealed time dependent effects of apoE-NT isoform binding to sLDLR. The results reveal a discerning method for study of the molecular basis of ligand interactions that likely influence receptor function in maintenance of whole body cholesterol homeostasis.

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A Receptor-Mediated Pathway for Cholesterol Homeostasis

Cited by 5,174 publications

References 146 publications

A novel method for determining functional LDL receptor activity in familial hypercholesterolemia: Application of the CD3/CD28 assay in lymphocytes

A novel method for determining functional LDL receptor activity in familial hypercholesterolemia: Application of the CD3/CD28 assay in lymphocytes

Molecular Characterization of Iranian Patients with Possible Familial Hypercholesterolemia

Apolipoprotein E isoform-specific binding to the low-density lipoprotein receptor

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