Aminopeptidase A

Ferreira, Anderson J.; Raizada, Mohan K.

doi:10.1161/hypertensionaha.107.109561

Cited by 9 publications

(2 citation statements)

References 11 publications

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“…we showed that APA cleaved brain AngII to generate angiotensin III (AngIII) in vivo and that AngIII exerts a tonic stimulatory action on the control of blood pressure (BP) in hypertensive animals [6][7][8]. The inhibition of brain APA with EC33 normalizes BP in alert hypertensive rats, suggesting that brain APA constitutes an interesting candidate target for the treatment of hypertension (HTN) [9]. This led us to develop an orally active APA inhibitor prodrug of EC33, RB150 renamed firibastat by World Health Organization, which blocks brain APA activity and AngIII formation and decreases BP in hypertensive rats [8,10,11].…”

Section: Introductionmentioning

confidence: 99%

Structural insight into the catalytic mechanism and inhibitor binding of aminopeptidase A

Couvineau

Almeida

Leroux

et al. 2020

Biochemical Journal

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Aminopeptidase A (APA) is a membrane-bound monozinc aminopeptidase. In the brain, APA generates angiotensin III which exerts a tonic stimulatory effect on the control of blood pressure in hypertensive animals. The oral administration of RB150 renamed firibastat by WHO, an APA inhibitor prodrug, targeting only the S1 subsite, decreases blood pressure in hypertensive patients from various ethnic origins. To identify new families of potent and selective APA inhibitors, we explored the organization of the APA active site, especially the S2’ subsite. By molecular modeling, docking, molecular dynamics simulations and site-directed mutagenesis, we revealed that Arg368 and Arg386, in the S2’ subsite of human APA established various types of interactions in major part with the P2’residue but also with the P1' residue of APA inhibitors, required for their nanomolar inhibitory potency. We also demonstrated an important role for Arg368 in APA catalysis, in maintaining the structural integrity of the GAMEN motif, a conserved sequence involved in exopeptidase specificity and optimal positioning of the substrate in monozinc aminopeptidases. This arginine together with the GAMEN motif are key players for the catalytic mechanism of these enzymes.

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Section: Introductionmentioning

confidence: 99%

Structural insight into the catalytic mechanism and inhibitor binding of aminopeptidase A

Couvineau

Almeida

Leroux

et al. 2020

Biochemical Journal

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“…Studies with specific and selective APA inhibitors (8) have shown that APA cleaved the N-terminal aspartyl residue of brain angiotensin II to generate angiotensin III in vivo (9) and that brain angiotensin III exerts a tonic stimulatory action on the control of blood pressure in hypertensive animals (10,11). Thus, the inhibition of brain APA with specific and selective inhibitors normalizes blood pressure in conscious spontaneously hypertensive rats or hypertensive deoxycorticosterone acetate salt rats (10 -12), suggesting that brain APA constitutes an interesting candidate target for the treatment of certain forms of hypertension (13)(14)(15). This justifies the development of potent and selective APA inhibitors crossing the blood-brain barrier after oral administration for use as centrally acting antihypertensive agents.…”

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confidence: 99%

Identification of Threonine 348 as a Residue Involved in Aminopeptidase A Substrate Specificity

Claperon

Banegas

Iturrioz

et al. 2009

Journal of Biological Chemistry

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Aminopeptidase A (APA; EC 3.4.11.7) is a membrane-bound zinc metalloprotease cleaving in the brain the N-terminal aspartyl residue of angiotensin II to generate angiotensin III, which exerts a tonic stimulatory effect on the central control of blood pressure in hypertensive animals. We docked the specific APA inhibitor, glutamate phosphonate, in the three-dimensional model of the mouse APA ectodomain in the presence of Ca 2؉ . In the S1 subsite of this model, the Ca 2؉ atom was coordinated with Asp-213, Asp-218,y and Glu-215 and three water molecules, one of which formed a hydrogen bond with the carboxylate side chain of the inhibitor. We report here that the carboxylate side chain of glutamate phosphonate also formed a hydrogen bond with the alcohol side chain of Thr-348. Mutagenic replacement of Thr-348 with an aspartate, tyrosine, or serine residue led to a modification of the hydrolysis velocity, with no change in the affinity of the recombinant enzymes for the substrate GluNA, either in the absence or presence of Ca 2؉ . In the absence of Ca 2؉ , the mutations modified the substrate specificity of APA, which was nevertheless restored by the addition of Ca 2؉ . An analysis of three-dimensional models of the corresponding Thr-348 mutants revealed that the interaction between this residue and the inhibitor was abolished or disturbed, leading to a change in the position of the inhibitor in the active site. These findings demonstrate a key role of Thr-348 in substrate specificity of APA for N-terminal acidic amino acids by insuring the optimal positioning of the substrate during catalysis.Aminopeptidase A (APA; EC 3.4.11.7) 3 is a 160-kDa homodimeric type II membrane-bound monozinc aminopeptidase also activated by Ca 2ϩ (1, 2). It specifically cleaves the N-terminal glutamyl or aspartyl residue from peptide substrates, such as angiotensin in vitro (3,4). APA is strongly expressed in many tissues, including the brush border of intestinal and renal epithelial cells, and in the vascular endothelium (5). APA has also been detected in several brain nuclei involved in controlling body fluid homeostasis and cardiovascular function (6, 7). Studies with specific and selective APA inhibitors (8) have shown that APA cleaved the N-terminal aspartyl residue of brain angiotensin II to generate angiotensin III in vivo (9) and that brain angiotensin III exerts a tonic stimulatory action on the control of blood pressure in hypertensive animals (10, 11). Thus, the inhibition of brain APA with specific and selective inhibitors normalizes blood pressure in conscious spontaneously hypertensive rats or hypertensive deoxycorticosterone acetate salt rats (10 -12), suggesting that brain APA constitutes an interesting candidate target for the treatment of certain forms of hypertension (13-15). This justifies the development of potent and selective APA inhibitors crossing the blood-brain barrier after oral administration for use as centrally acting antihypertensive agents. To achieve this goal, the study of the organization of the APA ac...

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Vitamin D and the renin–angiotensin system

2024

Feldman and Pike' S Vitamin D

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Aminopeptidase A

Cited by 9 publications

References 11 publications

Structural insight into the catalytic mechanism and inhibitor binding of aminopeptidase A

Structural insight into the catalytic mechanism and inhibitor binding of aminopeptidase A

Identification of Threonine 348 as a Residue Involved in Aminopeptidase A Substrate Specificity

Vitamin D and the renin–angiotensin system

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