Annexin A2 Reduces PCSK9 Protein Levels via a Translational Mechanism and Interacts with the M1 and M2 Domains of PCSK9

Ly, Kévin; Saavedra, Yascara Grisel Luna; Canuel, Maryssa; Routhier, Sophie; Desjardins, Roxane; Hamelin, Josée; Mayne, Janice; Lazure, Claude; Seidah, Nabil G.; Day, Robert

doi:10.1074/jbc.m113.541094

Cited by 46 publications

(43 citation statements)

References 54 publications

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“…The expression and secretion of both constructs (ϳ30 -35 kDa) were confirmed by Western blotting (Fig. 5, 1st and 4th lanes), as described previously (19,24). Only hPCSK9⌬CHRD-V5 coimmunoprecipitated with GPC3 by a 4.0-fold enrichment ratio compared with normal IgG (Fig.…”

Section: Validation Of Protein-proteinmentioning

confidence: 97%

“…Hence, we used a second approach to study the impact of the six binding candidates on LDL uptake in HepG2 cells by down-regulating their respective expression levels. We have previously assessed the role of AnxA2 and PCSK9 in LDL-cholesterol uptake in Huh7 and HepG2 cells by generating stable knockdown using lentivirus delivery of shRNAs (24). Herein, we generated the stable mRNA knockdown of GPC3, PLTP, MATN3, TFPI, FGL1, and PAI-1 in HepG2 cells using the same strategy in comparison with a stable HepG2 cell line expressing a nonspecific shRNA non-target (shNT) as a control (Table 1).…”

Section: Down-regulation Of Gpc3 and Pltp Exhibits Ldl Uptakementioning

confidence: 99%

“…However, it is also possible that other proteins could interact with the PCSK9 CHRD and act as a co-receptor for LDLR trafficking. Several studies have demonstrated that such interaction occurs extracellularly with the extrahepatic protein annexin A2 (AnxA2) acting as an endogenous inhibitor of PCSK9 via its ability to bind the CHRD and thus prevent its interaction with the LDLR (22)(23)(24).…”

mentioning

confidence: 99%

“…(13,(22)(23)(24)(25)(26)(27), several questions need to be addressed, including the mechanism leading the cell-surface PCSK9-LDLR complex to lysosomal degradation. Herein, we sought to identify novel extracellular interaction partners that could participate and regulate the PCSK9-LDLR complex formation and hence LDLR degradation.…”

mentioning

confidence: 99%

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An Unbiased Mass Spectrometry Approach Identifies Glypican-3 as an Interactor of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and Low Density Lipoprotein Receptor (LDLR) in Hepatocellular Carcinoma Cells

Essalmani

Desjardins

et al. 2016

Journal of Biological Chemistry

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Edited by Xiao-Fan WangThe mechanism of LDL receptor (LDLR) degradation mediated by the proprotein convertase subtilisin/kexin type 9 (PCSK9) has been extensively studied; however, many steps within this process remain unclear and still require characterization. Recent studies have shown that PCSK9 lacking its Cys/ His-rich domain can still promote LDLR internalization, but the complex does not reach the lysosome suggesting the presence of an additional interaction partner(s). In this study we carried out an unbiased screening approach to identify PCSK9-interacting proteins in the HepG2 cells' secretome using co-immunoprecipitation combined with mass spectrometry analyses. Several interacting proteins were identified, including glypican-3 (GPC3), phospholipid transfer protein, matrilin-3, tissue factor pathway inhibitor, fibrinogen-like 1, and plasminogen activator inhibitor-1. We then validated these interactions by co-immunoprecipitation and Western blotting. Furthermore, functional validation was examined by silencing each candidate protein in HepG2 cells using short hairpin RNAs to determine their effect on LDL uptake and LDLR levels. Only GPC3 and phospholipid transfer protein silencing in HepG2 cells significantly increased LDL uptake in these cells and displayed higher total LDLR protein levels compared with control cells. Moreover, our study provides the first evidence that GPC3 can modulate the PCSK9 extracellular activity as a competitive binding partner to the LDLR in HepG2 cells.

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Section: Validation Of Protein-proteinmentioning

confidence: 97%

Section: Down-regulation Of Gpc3 and Pltp Exhibits Ldl Uptakementioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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An Unbiased Mass Spectrometry Approach Identifies Glypican-3 as an Interactor of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and Low Density Lipoprotein Receptor (LDLR) in Hepatocellular Carcinoma Cells

Essalmani

Desjardins

et al. 2016

Journal of Biological Chemistry

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“…Extracellular AnxA2 was shown to also interact with the proprotein convertase subtilisin/kexin-type 9 (PCSK9), thus interfering with PCSK9-mediated degradation of the hepatic low-density lipoprotein receptor (LDLR) [85][86][87] In search of the molecular mechanism underlying the stimulatory effects of AnxA2 on human osteoclast formation [88,89], a novel type I membrane protein was identified as a putative AnxA2 receptor [90]. A recent study linked a single nucleotide polymorphism (SNP) in the AnxA2 gene (rs7170178) to osteonecrosis in sickle cell patients.…”

Section: Extracellular Functions-annexins As Ligands Of Defined Inflamentioning

confidence: 99%

Annexins in Translational Research: HIDDEN Treasures to Be Found

Schloer¹,

Pajonczyk²,

Rescher³

2018

Preprint

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The vertebrate annexin superfamily (AnxA) consists of 12 members of a calcium (Ca 2+ ) and phospholipid binding protein family which share a high structural homology. In keeping with this hallmark feature, annexins have been implicated in the Ca 2+ -controlled regulation of a broad range of membrane events. In this review, we identify and discuss several themes of annexin actions that hold a potential therapeutic value, namely the regulation of the immune response and the control of tissue homeostasis, and that repeatedly surface in the annexin activity profile. Our aim is to identify and discuss those annexin properties which might be exploited from a translational science and specifically clinical point of view.

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PCSK9 inhibitors as safer therapeutics for atherosclerotic cardiovascular disease (ASCVD): Pharmacophore design and molecular dynamics analysis

Kehinde,

Akawa,

Adewumi

et al. 2024

J of Cellular Biochemistry

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Cardiovascular disorders are still challenging and are among the deadly diseases. As a major risk factor for atherosclerotic cardiovascular disease, dyslipidemia, and high low‐density lipoprotein cholesterol in particular, can be prevented primary and secondary by lipid‐lowering medications. Therefore, insights are still needed into designing new drugs with minimal side effects. Proprotein convertase subtilisin/kexin 9 (PCSK9) enzyme catalyses protein‐protein interactions with low‐density lipoprotein, making it a critical target for designing promising inhibitors compared to statins. Therefore, we screened for potential compounds using a redesigned PCSK9 conformational behaviour to search for a significantly extensive chemical library and investigated the inhibitory mechanisms of the final compounds using integrated computational methods, from ligand essential functional group screening to all‐atoms MD simulations and MMGBSA‐based binding free energy. The inhibitory mechanisms of the screened compounds compared with the standard inhibitor. K31 and K34 molecules showed stronger interactions for PCSK9, having binding energy (kcal/mol) of −33.39 and −63.51, respectively, against −27.97 of control. The final molecules showed suitable drug‐likeness, non‐mutagenesis, permeability, and high solubility values. The C‐α atoms root mean square deviation and root mean square fluctuation of the bound‐PCSK9 complexes showed stable and lower fluctuations compared to apo PCSK9. The findings present a model that unravels the mechanism by which the final molecules proposedly inhibit the PCSK9 function and could further improve the design of novel drugs against cardiovascular diseases.

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Annexin A2 Reduces PCSK9 Protein Levels via a Translational Mechanism and Interacts with the M1 and M2 Domains of PCSK9

Cited by 46 publications

References 54 publications

An Unbiased Mass Spectrometry Approach Identifies Glypican-3 as an Interactor of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and Low Density Lipoprotein Receptor (LDLR) in Hepatocellular Carcinoma Cells

An Unbiased Mass Spectrometry Approach Identifies Glypican-3 as an Interactor of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and Low Density Lipoprotein Receptor (LDLR) in Hepatocellular Carcinoma Cells

Annexins in Translational Research: HIDDEN Treasures to Be Found

PCSK9 inhibitors as safer therapeutics for atherosclerotic cardiovascular disease (ASCVD): Pharmacophore design and molecular dynamics analysis

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