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A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles
Proprotein convertase subtilisin/kexin type-9 (PCSK9) is a ligand of low-density lipoprotein (LDL) receptor (LDLR) that promotes LDLR degradation in late endosomes/lysosomes. In human plasma, 30–40% of PCSK9 is bound to LDL particles; however, the physiological significance of this interaction remai...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7039556/ https://www.ncbi.nlm.nih.gov/pubmed/31949048 http://dx.doi.org/10.1074/jbc.RA119.010221 |
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author | Sarkar, Samantha K. Foo, Alexander C. Y. Matyas, Angela Asikhia, Ikhuosho Kosenko, Tanja Goto, Natalie K. Vergara-Jaque, Ariela Lagace, Thomas A. |
author_facet | Sarkar, Samantha K. Foo, Alexander C. Y. Matyas, Angela Asikhia, Ikhuosho Kosenko, Tanja Goto, Natalie K. Vergara-Jaque, Ariela Lagace, Thomas A. |
author_sort | Sarkar, Samantha K. |
collection | PubMed |
description | Proprotein convertase subtilisin/kexin type-9 (PCSK9) is a ligand of low-density lipoprotein (LDL) receptor (LDLR) that promotes LDLR degradation in late endosomes/lysosomes. In human plasma, 30–40% of PCSK9 is bound to LDL particles; however, the physiological significance of this interaction remains unknown. LDL binding in vitro requires a disordered N-terminal region in PCSK9's prodomain. Here, we report that peptides corresponding to a predicted amphipathic α-helix in the prodomain N terminus adopt helical structure in a membrane-mimetic environment. This effect was greatly enhanced by an R46L substitution representing an atheroprotective PCSK9 loss-of-function mutation. A helix-disrupting proline substitution within the putative α-helical motif in full-length PCSK9 lowered LDL binding affinity >5-fold. Modeling studies suggested that the transient α-helix aligns multiple polar residues to interact with positively charged residues in the C-terminal domain. Gain-of-function PCSK9 mutations associated with familial hypercholesterolemia (FH) and clustered at the predicted interdomain interface (R469W, R496W, and F515L) inhibited LDL binding, which was completely abolished in the case of the R496W variant. These findings shed light on allosteric conformational changes in PCSK9 required for high-affinity binding to LDL particles. Moreover, the initial identification of FH-associated mutations that diminish PCSK9's ability to bind LDL reported here supports the notion that PCSK9-LDL association in the circulation inhibits PCSK9 activity. |
format | Online Article Text |
id | pubmed-7039556 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-70395562020-03-04 A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles Sarkar, Samantha K. Foo, Alexander C. Y. Matyas, Angela Asikhia, Ikhuosho Kosenko, Tanja Goto, Natalie K. Vergara-Jaque, Ariela Lagace, Thomas A. J Biol Chem Lipids Proprotein convertase subtilisin/kexin type-9 (PCSK9) is a ligand of low-density lipoprotein (LDL) receptor (LDLR) that promotes LDLR degradation in late endosomes/lysosomes. In human plasma, 30–40% of PCSK9 is bound to LDL particles; however, the physiological significance of this interaction remains unknown. LDL binding in vitro requires a disordered N-terminal region in PCSK9's prodomain. Here, we report that peptides corresponding to a predicted amphipathic α-helix in the prodomain N terminus adopt helical structure in a membrane-mimetic environment. This effect was greatly enhanced by an R46L substitution representing an atheroprotective PCSK9 loss-of-function mutation. A helix-disrupting proline substitution within the putative α-helical motif in full-length PCSK9 lowered LDL binding affinity >5-fold. Modeling studies suggested that the transient α-helix aligns multiple polar residues to interact with positively charged residues in the C-terminal domain. Gain-of-function PCSK9 mutations associated with familial hypercholesterolemia (FH) and clustered at the predicted interdomain interface (R469W, R496W, and F515L) inhibited LDL binding, which was completely abolished in the case of the R496W variant. These findings shed light on allosteric conformational changes in PCSK9 required for high-affinity binding to LDL particles. Moreover, the initial identification of FH-associated mutations that diminish PCSK9's ability to bind LDL reported here supports the notion that PCSK9-LDL association in the circulation inhibits PCSK9 activity. American Society for Biochemistry and Molecular Biology 2020-02-21 2020-01-16 /pmc/articles/PMC7039556/ /pubmed/31949048 http://dx.doi.org/10.1074/jbc.RA119.010221 Text en © 2020 Sarkar et al. Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) . |
spellingShingle | Lipids Sarkar, Samantha K. Foo, Alexander C. Y. Matyas, Angela Asikhia, Ikhuosho Kosenko, Tanja Goto, Natalie K. Vergara-Jaque, Ariela Lagace, Thomas A. A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles |
title | A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles |
title_full | A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles |
title_fullStr | A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles |
title_full_unstemmed | A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles |
title_short | A transient amphipathic helix in the prodomain of PCSK9 facilitates binding to low-density lipoprotein particles |
title_sort | transient amphipathic helix in the prodomain of pcsk9 facilitates binding to low-density lipoprotein particles |
topic | Lipids |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7039556/ https://www.ncbi.nlm.nih.gov/pubmed/31949048 http://dx.doi.org/10.1074/jbc.RA119.010221 |
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