The Arg499His gain-of-function mutation in the C-terminal domain of PCSK9.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: Ireland NLM ID: 0242543 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-1484 (Electronic) Linking ISSN: 00219150 NLM ISO Abbreviation: Atherosclerosis Subsets: MEDLINE
    • Publication Information:
      Publication: Limerick : Elsevier
      Original Publication: Amsterdam, Elsevier.
    • Subject Terms:
      Gain of Function Mutation*; Arginine/*chemistry ; Histidine/*chemistry ; Proprotein Convertase 9/*genetics; Cell Membrane/metabolism ; Child ; Culture Media ; Family Health ; Female ; HEK293 Cells ; Hep G2 Cells ; Heterozygote ; Humans ; Italy ; Middle Aged ; Pedigree ; Protein Domains ; Receptors, LDL/metabolism ; Spain
    • Abstract:
      Background and Aims: Familial hypercholesterolemia (FH) is a monogenic disease characterized by high levels of low-density lipoprotein cholesterol and premature atherosclerotic cardiovascular disease. FH is caused by loss of function mutations in genes encoding LDL receptor (LDLR), and Apolipoprotein B (APOB) or gain of function (GOF) mutations in proprotein convertase subtilisin/kexin type 9 (PCSK9). In this study, we identified a novel variant in PCSK9, p.(Arg499His), located in the C-terminal domain, in two unrelated FH patients from Spain and Italy.
      Methods: We studied familial segregation and determined variant activity in vitro.
      Results: We determined PCSK9 expression, secretion and activity of the variant in transfected HEK293 cells; extracellular activity of the recombinant p.(Arg499His) PCSK9 variant in HEK 293 and HepG2 cells; PCSK9 affinity to the LDL receptor at neutral and acidic pH; the mechanism of action of the p.(Arg499His) PCSK9 variant by co-transfection with a soluble construct of the LDL receptor and by determining total PCSK9 intracellular accumulation when endosomal acidification is impaired and when an excess of soluble LDLr is present in the culture medium. Our results show high LDL-C concentrations and FH phenotype in p.(Arg499His) carriers. In vitro functional characterization shows that p.(Arg499His) PCSK9 variant causes a reduction in LDLr expression and LDL uptake. An intracellular activity for this variant is also shown when blocking the activity of secreted PCSK9 and by inhibiting endosomal acidification.
      Conclusions: We demonstrated that p.(Arg499His) PCSK9 variant causes a direct intracellular degradation of LDLr therefore causing FH by reducing LDLr availability.
      (Copyright © 2019 Elsevier B.V. All rights reserved.)
    • Contributed Indexing:
      Keywords: Cholesterol; Dyslipemia; Familial hypercholesterolemia; Functional assay; Gene expression; Genetics; Lipids; Mutations
    • Accession Number:
      0 (Culture Media)
      0 (LDLR protein, human)
      0 (Receptors, LDL)
      4QD397987E (Histidine)
      94ZLA3W45F (Arginine)
      EC 3.4.21.- (PCSK9 protein, human)
      EC 3.4.21.- (Proprotein Convertase 9)
    • Publication Date:
      Date Created: 20190914 Date Completed: 20200814 Latest Revision: 20200814
    • Publication Date:
      20240829
    • Accession Number:
      10.1016/j.atherosclerosis.2019.08.020
    • Accession Number:
      31518966