Effects of PCSK9 missense variants on molecular conformation and biological activity in transfected HEK293FT cells
Gene
; 851(146979)Oct. 2022.
Article
in English
| CONASS, Sec. Est. Saúde SP, SESSP-IDPCPROD, Sec. Est. Saúde SP
| ID: biblio-1400683
Responsible library:
BR79.1
ABSTRACT
ABSTRACT 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.
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Collection:
National databases
/
Brazil
Database:
CONASS
/
Sec. Est. Saúde SP
/
SESSP-IDPCPROD
Main subject:
Mutation, Missense
/
Hyperlipoproteinemia Type II
/
Molecular Conformation
Limits:
Humans
Language:
English
Journal:
Gene
Year:
2022
Document type:
Article
Institution/Affiliation country:
Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of Sao Paulo/BR
/
Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo/BR
/
Department of Internal Medicine VIII, University Hospitals Tübingen/DE
/
Laboratory of Genetics and Molecular Cardiology, Heart Institute, University of São Paulo/BR
/
Laboratory of Molecular Research in Cardiology, Institute Dante Pazzanese of Cardiology/BR
/
Medical Clinic Division, Institute Dante Pazzanese of Cardiology/BR