Elucidation of the Structure–Activity Relationships of Apelin: Influence of Unnatural Amino Acids on Binding, Signaling, and Plasma Stability

Murza, Alexandre; Parent, Alexandre J.; Besserer-Offroy, Élie; Tremblay, Hugo; Karadereye, Félix; Beaudet, Nicolas; Leduc, Richard; Sarret, Philippe; Marsault, Éric

doi:10.1002/cmdc.201100492

Cited by 67 publications

(126 citation statements)

References 46 publications

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“…We modified, synthesized, and purified 2 novel apelin analogues, NleAibBrF-pyr-apelin 13 and NleAibBrF-apelin 17, using a previously described method. 8 Using structure-activity relationships conducted on pyr-apelin 13, 8,34 we made multiple novel single amino acid substitutions combined into the same peptide with the aim of masking the susceptible C-terminal amide bond from proteolytic cleavage. These analogues were purified using HPLC, and high-resolution mass spectrometry and nuclear magnetic resonance were used to confirm the sequence of the synthesized analogues ( Figure 6A and 6B).…”

Section: Design and Synthesis Of Apelin Analogues Resistant To Ace2 Dmentioning

confidence: 99%

Angiotensin-Converting Enzyme 2 Metabolizes and Partially Inactivates Pyr-Apelin-13 and Apelin-17

et al. 2016

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Abstract-Apelin peptides mediate beneficial effects on the cardiovascular system and are being targeted as potential new drugs. However, apelin peptides have extremely short biological half-lives, and improved understanding of apelin peptide metabolism may lead to the discovery of biologically stable analogues with therapeutic potential. We examined the ability of angiotensin-converting enzyme 2 (ACE2) to cleave and inactivate pyr-apelin 13 and apelin 17, the dominant apelin peptides. Computer-assisted modeling shows a conserved binding of pyr-apelin 13 and apelin 17 to the ACE2 catalytic site. In ACE2 knockout mice, hypotensive action of pyr-apelin 13 and apelin 17 was potentiated, with a corresponding greater elevation in plasma apelin levels. Similarly, pharmacological inhibition of ACE2 potentiated the vasodepressor action of apelin peptides. Biochemical analysis confirmed that recombinant human ACE2 can cleave pyr-apelin 13 and apelin 17 efficiently, and apelin peptides are degraded slower in ACE2-deficient plasma. The biological relevance of ACE2-mediated proteolytic processing of apelin peptides was further supported by the reduced potency of pyr-apelin 12 and apelin 16 on the activation of signaling pathways and nitric oxide production from endothelial cells. Importantly, although pyr-apelin 13 and apelin 17 rescued contractile function in a myocardial ischemia-reperfusion model, ACE2 cleavage products, pyr-apelin 12 and 16, were devoid of these cardioprotective effects. We designed and synthesized active apelin analogues that were resistant to ACE2-mediated degradation, thereby confirming that stable apelin analogues can be designed as potential drugs. We conclude that ACE2 represents a major negative regulator of apelin action in the vasculature and heart. residue and the enzymatic processes involved in its removal from the native apelin peptide remains poorly defined. Using loss-of-function and gain-of-function strategies, we here define a critical role of angiotensin-converting enzyme 2 (ACE2) in the proteolytic cleavage of the C-terminal phenylalanine residue in pyr-apelin 13 and apelin 17. Importantly, this degradative site can be modified to produce relatively stable apelin analogues as potential therapeutic agents. Methods In Silico Modeling of Apelin Peptide Binding to ACE2We selected the structure of human apo-ACE2 (PDB ID: 1R42) mainly because of its high resolution, appropriate R-factor, and errorless electron density map. 16 This structure was equilibrated in constant pressure-temperature condition (NVT, NPT) in the Groningen Machine for Chemical Simulations (GROMACS). 17 We performed knowledgebased docking using our understanding from ACE-substrate bound complex structure. For this purpose, we have used the Angiotensin II (Ang II)-bound ACE complex (PDB ID: 4APH) 18 and MLN-4760 inhibitor bound ACE2 complex (PDB ID: 1R4L). 16,[18][19][20] We modeled the ACE2-Ang II, ACE2-pyr-apelin 13, and ACE2-apelin 17 complexes using these 2 above mentioned reference structures as a template. We built th...

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Section: Design and Synthesis Of Apelin Analogues Resistant To Ace2 Dmentioning

confidence: 99%

Angiotensin-Converting Enzyme 2 Metabolizes and Partially Inactivates Pyr-Apelin-13 and Apelin-17

et al. 2016

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“…Compellingly, the conserved structural features of the apelin isoforms have been directly employed in design of peptide-based AR antagonists (Macaluso and Glen 2010;Macaluso et al 2011), and agonists (Murza et al 2012). Despite the clear importance of the apelin C-terminal dodecapeptide region, the presence of multiple apelin isoforms in the body suggests an evolutionary purpose.…”

Section: Apelin-induced Signallingmentioning

confidence: 99%

The apelin receptor: physiology, pathology, cell signalling, and ligand modulation of a peptide-activated class A GPCR

Chapman

Dupré

Rainey

2014

Biochem. Cell Biol.

171

135

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The apelin receptor (AR or APJ) is a class A (rhodopsin-like) G-protein coupled receptor (GPCR) with wide distribution throughout the human body. Activation of the AR by its cognate peptide ligand, apelin, induces diverse physiological effects including vasoconstriction and dilation; strengthening of heart muscle contractility; angiogenesis; and, regulation of energy metabolism and fluid homeostasis. Recently, another endogenous peptidic activator of the AR, Toddler/ ELABELA, was identified as having a crucial role in zebrafish embryonic development. The AR is also implicated in pathologies including cardiovascular disease, diabetes, obesity and cancer, making it a promising therapeutic target. Despite its established importance, the precise roles of AR signalling remain poorly understood. Moreover, little is known about mechanisms of peptide-AR activation. Additional complexity arises from modulation of the AR by two endogenous peptide ligands, both with multiple bioactive isoforms of variable length and distribution. The various apelin and Toddler/ELABELA isoforms may also produce distinct cellular effects. Further complexity arises through formation of functionally distinct heterodimers between the AR and other GPCRs. This minireview outlines key (patho)physiological actions of the AR, addresses what is known about signal transduction downstream of AR activation, and concludes by discussing unique properties of the endogenous peptidic ligands of the AR.

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“…Apelin peptides are readily metabolized in vivo with a half-life ( t 1/2 ) of ∼5 min. 10 Past studies have implicated angiotensin converting enzyme 2 (ACE2) in cleavage of the C-terminal phenylalanine residue of apelin-13. 11 However, data also indicate that absence or alteration of this residue does not lead to complete inactivation of apelin peptides in vitro.…”

Section: Endogenous Ligands Of the Apelin Receptormentioning

confidence: 99%

Regulation of the Apelinergic System and Its Potential in Cardiovascular Disease: Peptides and Small Molecules as Tools for Discovery

et al. 2015

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Apelin peptides and the apelin receptor represent a relatively new therapeutic axis for the potential treatment of cardiovascular disease. Several reports suggest apelin receptor activation with apelin peptides results in cardioprotection as noted through positive ionotropy, angiogenesis, reduction of mean arterial blood pressure, and apoptosis. Considering the potential therapeutic benefit attainable through modulation of the apelinergic system, research is expanding to develop novel therapies that limit the inherent rapid degradation of endogenous apelin peptides and produce metabolically stable small molecule agonists and antagonists to more rigorously interrogate the apelin receptor system. This review details the structure–activity relationships for chemically modified apelin peptides and recent disclosures of small molecule agonists and antagonists and summarizes the peer reviewed and patented literature. Development of metabolically stable ligands of apelin receptor and their effects in various models over the coming years will hopefully lead to establishment of this receptor as a validated target for cardiovascular indications.

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Elucidation of the Structure–Activity Relationships of Apelin: Influence of Unnatural Amino Acids on Binding, Signaling, and Plasma Stability

Cited by 67 publications

References 46 publications

Angiotensin-Converting Enzyme 2 Metabolizes and Partially Inactivates Pyr-Apelin-13 and Apelin-17

Angiotensin-Converting Enzyme 2 Metabolizes and Partially Inactivates Pyr-Apelin-13 and Apelin-17

The apelin receptor: physiology, pathology, cell signalling, and ligand modulation of a peptide-activated class A GPCR

Regulation of the Apelinergic System and Its Potential in Cardiovascular Disease: Peptides and Small Molecules as Tools for Discovery

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