Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject
Familial hypercholesterolaemia (FH) is an autosomal dominant dyslipidaemia, characterised by elevated LDL cholesterol (LDL-C) levels in the blood. Three main genes are involved in FH diagnosis: LDL receptor (LDLr), Apolipoprotein B (APOB) and Protein convertase subtilisin/kexin type 9 (PCSK9) with g...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-02-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/4/3330 |
| _version_ | 1797620544653230080 |
|---|---|
| author | Asier Larrea-Sebal Chiara Trenti Shifa Jebari-Benslaiman Stefano Bertolini Sebastiano Calandra Emanuele A. Negri Efrem Bonelli Asier Benito-Vicente Leire Uraga-Gracianteparaluceta César Martín Tommaso Fasano |
| author_facet | Asier Larrea-Sebal Chiara Trenti Shifa Jebari-Benslaiman Stefano Bertolini Sebastiano Calandra Emanuele A. Negri Efrem Bonelli Asier Benito-Vicente Leire Uraga-Gracianteparaluceta César Martín Tommaso Fasano |
| author_sort | Asier Larrea-Sebal |
| collection | DOAJ |
| description | Familial hypercholesterolaemia (FH) is an autosomal dominant dyslipidaemia, characterised by elevated LDL cholesterol (LDL-C) levels in the blood. Three main genes are involved in FH diagnosis: LDL receptor (LDLr), Apolipoprotein B (APOB) and Protein convertase subtilisin/kexin type 9 (PCSK9) with genetic mutations that led to reduced plasma LDL-C clearance. To date, several PCSK9 gain-of-function (GOF) variants causing FH have been described based on their increased ability to degrade LDLr. On the other hand, mutations that reduce the activity of PCSK9 on LDLr degradation have been described as loss-of-function (LOF) variants. It is therefore important to functionally characterise PCSK9 variants in order to support the genetic diagnosis of FH. The aim of this work is to functionally characterise the p.(Arg160Gln) PCSK9 variant found in a subject suspected to have FH. Different techniques have been combined to determine efficiency of the autocatalytic cleavage, protein expression, effect of the variant on LDLr activity and affinity of the PCSK9 variant for the LDLr. Expression and processing of the p.(Arg160Gln) variant had a result similar to that of WT PCSK9. The effect of p.(Arg160Gln) PCSK9 on LDLr activity is lower than WT PCSK9, with higher values of LDL internalisation (13%) and p.(Arg160Gln) PCSK9 affinity for the LDLr is lower than WT, EC50 8.6 ± 0.8 and 25.9 ± 0.7, respectively. The p.(Arg160Gln) PCSK9 variant is a LOF PCSK9 whose loss of activity is caused by a displacement of the PCSK9 P’ helix, which reduces the stability of the LDLr-PCSK9 complex. |
| first_indexed | 2024-03-11T08:44:06Z |
| format | Article |
| id | doaj.art-83ca415e3111468c8d3293155e452740 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T08:44:06Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-83ca415e3111468c8d3293155e4527402023-11-16T20:58:28ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01244333010.3390/ijms24043330Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic SubjectAsier Larrea-Sebal0Chiara Trenti1Shifa Jebari-Benslaiman2Stefano Bertolini3Sebastiano Calandra4Emanuele A. Negri5Efrem Bonelli6Asier Benito-Vicente7Leire Uraga-Gracianteparaluceta8César Martín9Tommaso Fasano10Department of Biochemistry and Molecular Biology, Universidad del País Vasco UPV/EHU, 48080 Bilbao, SpainDepartment of Internal Medicine Unit, Azienda USL—IRCCS di Reggio Emilia, 42122 Reggio Emilia, ItalyDepartment of Biochemistry and Molecular Biology, Universidad del País Vasco UPV/EHU, 48080 Bilbao, SpainDepartment of Internal Medicine, University of Genova, 16132 Genova, ItalyDepartment of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41125 Modena, ItalyDepartment of Internal Medicine Unit, Azienda USL—IRCCS di Reggio Emilia, 42122 Reggio Emilia, ItalyDepartment of Internal Medicine Unit, Azienda USL—IRCCS di Reggio Emilia, 42122 Reggio Emilia, ItalyDepartment of Biochemistry and Molecular Biology, Universidad del País Vasco UPV/EHU, 48080 Bilbao, SpainDepartment of Biochemistry and Molecular Biology, Universidad del País Vasco UPV/EHU, 48080 Bilbao, SpainDepartment of Biochemistry and Molecular Biology, Universidad del País Vasco UPV/EHU, 48080 Bilbao, SpainClinical Pathology Unit, AUSL Romagna, 47521 Cesena, ItalyFamilial hypercholesterolaemia (FH) is an autosomal dominant dyslipidaemia, characterised by elevated LDL cholesterol (LDL-C) levels in the blood. Three main genes are involved in FH diagnosis: LDL receptor (LDLr), Apolipoprotein B (APOB) and Protein convertase subtilisin/kexin type 9 (PCSK9) with genetic mutations that led to reduced plasma LDL-C clearance. To date, several PCSK9 gain-of-function (GOF) variants causing FH have been described based on their increased ability to degrade LDLr. On the other hand, mutations that reduce the activity of PCSK9 on LDLr degradation have been described as loss-of-function (LOF) variants. It is therefore important to functionally characterise PCSK9 variants in order to support the genetic diagnosis of FH. The aim of this work is to functionally characterise the p.(Arg160Gln) PCSK9 variant found in a subject suspected to have FH. Different techniques have been combined to determine efficiency of the autocatalytic cleavage, protein expression, effect of the variant on LDLr activity and affinity of the PCSK9 variant for the LDLr. Expression and processing of the p.(Arg160Gln) variant had a result similar to that of WT PCSK9. The effect of p.(Arg160Gln) PCSK9 on LDLr activity is lower than WT PCSK9, with higher values of LDL internalisation (13%) and p.(Arg160Gln) PCSK9 affinity for the LDLr is lower than WT, EC50 8.6 ± 0.8 and 25.9 ± 0.7, respectively. The p.(Arg160Gln) PCSK9 variant is a LOF PCSK9 whose loss of activity is caused by a displacement of the PCSK9 P’ helix, which reduces the stability of the LDLr-PCSK9 complex.https://www.mdpi.com/1422-0067/24/4/3330familial hypercholesterolemiaPCSK9GOFLOFcharacterisationactivity |
| spellingShingle | Asier Larrea-Sebal Chiara Trenti Shifa Jebari-Benslaiman Stefano Bertolini Sebastiano Calandra Emanuele A. Negri Efrem Bonelli Asier Benito-Vicente Leire Uraga-Gracianteparaluceta César Martín Tommaso Fasano Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject International Journal of Molecular Sciences familial hypercholesterolemia PCSK9 GOF LOF characterisation activity |
| title | Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject |
| title_full | Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject |
| title_fullStr | Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject |
| title_full_unstemmed | Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject |
| title_short | Functional Characterization of p.(Arg160Gln) PCSK9 Variant Accidentally Found in a Hypercholesterolemic Subject |
| title_sort | functional characterization of p arg160gln pcsk9 variant accidentally found in a hypercholesterolemic subject |
| topic | familial hypercholesterolemia PCSK9 GOF LOF characterisation activity |
| url | https://www.mdpi.com/1422-0067/24/4/3330 |
| work_keys_str_mv | AT asierlarreasebal functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT chiaratrenti functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT shifajebaribenslaiman functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT stefanobertolini functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT sebastianocalandra functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT emanueleanegri functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT efrembonelli functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT asierbenitovicente functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT leireuragagracianteparaluceta functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT cesarmartin functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject AT tommasofasano functionalcharacterizationofparg160glnpcsk9variantaccidentallyfoundinahypercholesterolemicsubject |