Role of the N-terminal Catalytic Domain of Angiotensin-converting Enzyme Investigated by Targeted Inactivation in Mice

Fuchs, Sébastien; Xiao, Hong; Cole, Justin M.; Adams, Jonathan; Frenzel, Kristen; Michaud, Annie; Zhao, Hui; Keshelava, George; Capecchi, Mario R.; Corvol, Pierre; Bernstein, Kenneth E.

doi:10.1074/jbc.m400149200

Cited by 95 publications

(96 citation statements)

References 35 publications

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“…As previously reported, we found that ACE was highly expressed in kidney and lung but absent in the liver (36). In the brain, we observed similar expression of ACE protein in the cerebral cortex, hippocampus, cerebellum, and basal ganglia/brainstem (Fig.…”

Section: Expression and Characterization Of Transfected Human Ace-tosupporting

confidence: 73%

Amyloid β-Protein Is Degraded by Cellular Angiotensin-converting Enzyme (ACE) and Elevated by an ACE Inhibitor

Hemming

Selkoe

2005

Journal of Biological Chemistry

295

211

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Human genetic data have associated angiotensin-converting enzyme (ACE) with Alzheimer disease (AD), and purified ACE has been reported to cleave synthetic amyloid ␤-protein (A␤) in vitro. Whether deficiency in ACE activity, arising from genetic alteration or pharmacological inhibition, can decrease A␤ degradation and allow A␤ accumulation in intact cells is unknown. We cloned ACE from human neuroblastoma cells and showed that it had posttranslational processing and enzymatic activity typical of the endogenous protease. Cellular expression of ACE promoted degradation of naturally secreted A␤40 and A␤42, leading to significant clearance of both species. Using site-directed mutagenesis, we determined that both active sites within ACE contribute to A␤ clearance, and an ACE construct bearing mutations in each catalytic domain had no effect on A␤ levels. Pharmacological inhibition of ACE with a widely prescribed drug, captopril, promoted the accumulation of cell-derived A␤ in the media of ␤-amyloid precursor-protein expressing cells. Together, these results show that ACE can lower the levels of secreted A␤ in living cells and that this effect is blocked by inhibiting the protease's activity with an ACE inhibitor. This work, combined with the genetic studies, supports the hypothesis that ACE may modulate the susceptibility to and progression of AD via degradation of A␤. Our data encourage further analyses of the ACE gene for disease association and raise the question of whether currently prescribed ACE inhibitors could elevate cerebral A␤ levels in humans. An early and pathogenically important feature of Alzheimer disease (AD)2 is the progressive accumulation and deposition of the amyloid ␤-protein (A␤) in brain regions serving memory and cognition. Biochemical, cell biological, animal modeling, genetic, and emerging clinical data all suggest that A␤ is an upstream initiator of the disease process and its associated neuropathology (1-4). Although no proven diseasemodifying treatments are currently available, recent efforts to treat AD have focused on both decreasing the production of A␤ and enhancing its clearance from the brain. One little studied approach to A␤ clearance is augmenting the degradation of the peptide by various proteases expressed in the brain. Thus far, the metalloproteases neprilysin (NEP) (5), insulin-degrading enzyme (IDE) (6), and the endothelin-converting enzymes 1 and 2 (7) have each been implicated as A␤-degrading proteases in the mammalian brain. The serine protease plasmin has been implicated in A␤ degradation in vitro (8), although genetic plasmin deficiency did not promote accumulation of murine A␤ in vivo (9). Supporting a role for therapeutic regulation of A␤-degrading proteases, the overexpression of IDE or NEP in a murine model of AD decreased cerebral A␤ levels and produced significant attenuation of A␤-associated neuropathology (10).Somatic angiotensin-converting enzyme (ACE) is a zinc metalloprotease containing two homologous regions, termed the N-and C-domains, each of which is prot...

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Section: Expression and Characterization Of Transfected Human Ace-tosupporting

confidence: 73%

Amyloid β-Protein Is Degraded by Cellular Angiotensin-converting Enzyme (ACE) and Elevated by an ACE Inhibitor

Hemming

Selkoe

2005

Journal of Biological Chemistry

295

211

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“…1, C and F), suggesting that each catalytic domain of ACE regulates the activity of the other, and both domains are required for normal substrate recognition and degradation. Mice with a selective inactivation of either the N-or C-domain of ACE were generated, and the C-domain was demonstrated to be the main site of angiotensin I cleavage (18,22), which is consistent with our in vitro finding. However, the role of the N-domain of ACE toward A␤42 to A␤40 conversion in vivo needs to be addressed.…”

Section: Discussionsupporting

confidence: 78%

Aβ42-to-Aβ40- and Angiotensin-converting Activities in Different Domains of Angiotensin-converting Enzyme

Zou

Maeda

Watanabe

et al. 2009

Journal of Biological Chemistry

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Amyloid ␤-protein 1-42 (A␤42) is believed to play a causative role in the development of Alzheimer disease (AD), although it is a minor part of A␤. In contrast, A␤40 is the predominant secreted form of A␤ and recent studies have suggested that A␤40 has neuroprotective effects and inhibits amyloid deposition. We have reported that angiotensin-converting enzyme (ACE) converts A␤42 to A␤40, and its inhibition enhances brain A␤42 deposition (Zou, K., Yamaguchi, H., Akatsu, H., Sakamoto, T., Ko, M., Mizoguchi, K., Gong, J. S., Yu, W., Yamamoto, T., Kosaka, K., Yanagisawa, K., and Michikawa, M. (2007) J. Neurosci. 27, 8628 -8635). ACE has two homologous domains, each having a functional active site. In the present study, we identified the domain of ACE, which is responsible for converting A␤42 to A␤40. Interestingly, A␤42-to-A␤40-converting activity is solely found in the N-domain of ACE and the angiotensinconverting activity is found predominantly in the C-domain of ACE. We also found that the N-linked glycosylation is essential for both A␤42-to-A␤40-and angiotensin-converting activities and that unglycosylated ACE rapidly degraded. The domainspecific converting activity of ACE suggests that ACE inhibitors could be designed to specifically target the angiotensin-converting C-domain, without inhibiting the A␤42-to-A␤40-converting activity of ACE or increasing neurotoxic A␤42. Angiotensin-converting enzyme (ACE)4 plays a key role in the renin-angiotensin system (RAS), which is involved in the long-term regulation of blood pressure and blood volume in the human body. Recent genetic, pathologic, and biochemical studies have associated ACE with onset of Alzheimer disease (AD) (1, 2). The I allele of the ACE gene, which results in a reduced serum ACE level, has been demonstrated to be associated with AD (3-5). Hypertension is a risk factor for AD and ACE inhibitors for treatment of hypertension were shown to be the only drug class among the antihypertensives to potentially be associated with a slight increased incidence of AD (adjusted hazard ratio 1.13) (6, 7). A mechanistic link between ACE and AD was suggested when ACE was shown to degrade A␤40 and A␤42 (8, 9). Overexpression of A␤40 in transgenic mice does not cause brain amyloid deposition, the major pathological hallmark of AD, whereas expression of A␤42 is shown to be essential for amyloid deposition (10, 11). In addition, A␤40 has an inhibitory effect on amyloid deposition in vitro and in vivo and has neuroprotective effects (12)(13)(14). These lines of evidence suggest that converting A␤42 to A␤40 may be a potential strategy for development of an AD therapy. In our previous study, we identified ACE as an A␤42-to-A␤40-converting (A␤-converting) enzyme and showed that ACE inhibitor enhances brain A␤42 deposition in transgenic mice (15). Clarifying the molecular base of ACE domain-specific enzymatic activity on A␤42 to A␤40 conversion, A␤ degradation, and angiotensin conversion emerges to be important for development of a strategy for hypertension and AD treatment.AC...

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“…26,27 We, therefore, asked whether MWF rats could exhibit changes in antifibrotic Ac-SDKP levels that may reflect altered Prep-dependent generation and/or ACE-mediated clearance of the peptide, and also, we asked whether they could be modulated as part of the renoprotective action of ACEi.…”

Section: Modulation Of Ac-sdkp Contributes To Reduction Of Interstitimentioning

confidence: 99%

MicroRNA-324-3p Promotes Renal Fibrosis and Is a Target of ACE Inhibition

Macconi

Tomasoni

Romagnani

et al. 2012

Journal of the American Society of Nephrology

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The contribution of microRNA (miRNA) to the pathogenesis of renal fibrosis is not well understood. Here, we investigated whether miRNA modulates the fibrotic process in Munich Wistar Fromter (MWF) rats, which develop spontaneous progressive nephropathy. We analyzed the expression profile of miRNA in microdissected glomeruli and found that miR-324-3p was the most upregulated. In situ hybridization localized miR-324-3p to glomerular podocytes, parietal cells of Bowman's capsule, and most abundantly, cortical tubules. A predicted target of miR-324-3p is prolyl endopeptidase (Prep), a serine peptidase involved in the metabolism of angiotensins and the synthesis of the antifibrotic peptide N-acetyl-serylaspartyl-lysyl-proline (Ac-SDKP). In cultured tubular cells, transient transfection with a miR-324-3p mimic reduced Prep protein and activity, validating Prep as a target of this miRNA. In MWF rats, upregulation of miR-324-3p associated with markedly reduced expression of Prep in both glomeruli and tubules, low urine Ac-SDKP, and increased deposition of collagen. ACE inhibition downregulated glomerular and tubular miR-324-3p, promoted renal Prep expression, increased plasma and urine Ac-SDKP, and attenuated renal fibrosis. In summary, these results suggest that dysregulation of the miR-324-3p/Prep pathway contributes to the development of fibrosis in progressive nephropathy. The renoprotective effects of ACE inhibitors may result, in part, from modulation of this pathway, suggesting that it may hold other potential therapeutic targets. 23: 149623: -150523: , 201223: . doi: 10.1681 Munich Wistar Fromter (MWF) is an inbred rat strain characterized by inborn nephron deficit, glomerular hypertrophy, and high urinary protein excretion. 1,2 Males develop hypertension and overt proteinuria with age. 3,4 Angiotensin II (Ang II) has a direct role in the pathogenesis of proteinuria and glomerular injury by decreasing the size selectivity of the glomerular capillary barrier against the filtration of plasma proteins 5,6 that is followed by development of glomerulosclerosis. 4 Treatment with an angiotensin-converting enzyme inhibitor (ACEi) protects MWF rats from kidney damage by both preserving and remodeling the glomerular barrier, thus inducing regression of proteinuria and glomerulosclerosis. 7,8 A large body of evidence in this model and other animal models and clinical trials confirms that Ang II is a potential therapeutic target for remission/regression of progressive glomerulopathies. 9,10 In advanced nephropathy, J Am Soc Nephrol

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Role of the N-terminal Catalytic Domain of Angiotensin-converting Enzyme Investigated by Targeted Inactivation in Mice

Cited by 95 publications

References 35 publications

Amyloid β-Protein Is Degraded by Cellular Angiotensin-converting Enzyme (ACE) and Elevated by an ACE Inhibitor

Amyloid β-Protein Is Degraded by Cellular Angiotensin-converting Enzyme (ACE) and Elevated by an ACE Inhibitor

Aβ42-to-Aβ40- and Angiotensin-converting Activities in Different Domains of Angiotensin-converting Enzyme

MicroRNA-324-3p Promotes Renal Fibrosis and Is a Target of ACE Inhibition

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