Effects of PCSK9 missense variants on molecular conformation and biological activity in transfected HEK293FT cells.
Familial hypercholesterolemia
In vitro functional study
Missense variants
Molecular modeling
PCSK9
Journal
Gene
ISSN: 1879-0038
Titre abrégé: Gene
Pays: Netherlands
ID NLM: 7706761
Informations de publication
Date de publication:
30 Jan 2023
30 Jan 2023
Historique:
received:
11
08
2022
revised:
30
09
2022
accepted:
12
10
2022
pubmed:
20
10
2022
medline:
30
11
2022
entrez:
19
10
2022
Statut:
ppublish
Résumé
PCSK9 gain-of-function (GOF) variants increase degradation of low-density lipoprotein receptor (LDLR) and are potentially associated with Familial Hypercholesterolemia (FH). This study aimed to explore the effects of PCSK9 missense variants on protein structure and interactions with LDLR using molecular modeling analyses and in vitro functional studies. Variants in FH-related genes were identified in a Brazilian FH cohort using an exon-target gene sequencing strategy. Eight PCSK9 missense variants in pro- [p.(E32K) and p.(E57K)], catalytic [p.(R237W), p.(P279T) and p.(A443T)], and C-terminal histidine-cysteine rich (CHR) [p.(R469W), p.(Q619P) and p.(R680Q)] domains were identified. Molecular dynamics analyses revealed that GOF variants p.(E32K) and p.(R469W) increased extreme motions in PCSK9 amino acid backbone fluctuations and affected Hbond and water bridge interactions between the pro-domain and CM1 region of the CHR domain. HEK293FT cells transfected with plasmids carrying p.(E32K) and p.(R469W) variants reduced LDLR expression (8.7 % and 14.8 %, respectively) compared to wild type (p < 0.05) but these GOF variants did not affect PCSK9 expression and secretion. The missense variants p.(P279T) and p.(Q619P) also reduced protein stability and altered Hbond interactions. In conclusion, PCSK9 p.(E32K), p.(R469W), p.(P279T) and p.(Q619P) variants disrupt intramolecular interactions that are essential for PCSK9 structural conformation and biological activity and may have a potential role in FH pathogenesis.
Identifiants
pubmed: 36261084
pii: S0378-1119(22)00799-5
doi: 10.1016/j.gene.2022.146979
pii:
doi:
Substances chimiques
PCSK9 protein, human
EC 3.4.21.-
Proprotein Convertase 9
EC 3.4.21.-
Receptors, LDL
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
146979Informations de copyright
Copyright © 2022 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mario Hiroyuki Hirata reports financial support was provided by State of Sao Paulo Research Foundation. Mario Hiroyuki Hirata reports financial support was provided by National Council for Scientific and Technological Development. Mario Hiroyuki Hirata reports financial support was provided by Coordination of Higher Education Personnel Improvement.