The N Domain of Human Angiotensin-I-converting Enzyme

Anthony, Colin; Corradi, H.R.; Schwager, S.L.U.; Redelinghuys, Pierre; Georgiadis, Dimitris; Dive, Vincent; Acharya, K. Ravi; Sturrock, Edward D.

doi:10.1074/jbc.m110.167866

Cited by 77 publications

(70 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, the determination of the RXP407 co-crystallized with the N-domain shows that, of all the unique amino acids present, only prominent contacts with the unique residues Tyr 369 and Arg 381 (by the P 2 aspartate residue) were exploited by the inhibitor [34]. This observation is consistent with a mutagenic study [35].…”

Section: Introductionsupporting

confidence: 67%

Fragment-based design for the development of N-domain-selective angiotensin-1-converting enzyme inhibitors

et al. 2013

View full text Add to dashboard Cite

ACE (angiotensin-1-converting enzyme) is a zinc metallopeptidase that plays a prominent role in blood pressure regulation and electrolyte homeostasis. ACE consists of two homologous domains that despite similarities of sequence and topology display differences in substrate processing and inhibitor binding. The design of inhibitors that selectively inhibit the N-domain (N-selective) could be useful in treating conditions of tissue injury and fibrosis due to build-up of N-domain-specific substrate Ac-SDKP (N-acetyl-Ser–Asp–Lys–Pro). Using a receptor-based SHOP (scaffold hopping) approach with N-selective inhibitor RXP407, a shortlist of scaffolds that consisted of modified RXP407 backbones with novel chemotypes was generated. These scaffolds were selected on the basis of enhanced predicted interaction energies with N-domain residues that differed from their C-domain counterparts. One scaffold was synthesized and inhibitory binding tested using a fluorogenic ACE assay. A molecule incorporating a tetrazole moiety in the P2 position (compound 33RE) displayed potent inhibition (Ki=11.21±0.74 nM) and was 927-fold more selective for the N-domain than the C-domain. A crystal structure of compound 33RE in complex with the N-domain revealed its mode of binding through aromatic stacking with His388 and a direct hydrogen bond with the hydroxy group of the N-domain specific Tyr369. This work further elucidates the molecular basis for N-domainselective inhibition and assists in the design of novel N-selective ACE inhibitors that could be employed in treatment of fibrosis disorders.

show abstract

Section: Introductionsupporting

confidence: 67%

Fragment-based design for the development of N-domain-selective angiotensin-1-converting enzyme inhibitors

et al. 2013

View full text Add to dashboard Cite

show abstract

“…In the nACE structures, sections of this region often show flexibility with increased temperature factors (B‐factors) and poor electron density. This is typically more apparent in one chain of the asymmetric unit, and this region has a small, up to 3Ẳ, shift between the two chains 20. Both the sampatrilat and samAsp nACE structures exhibit this flexibility in the hinge region.…”

Section: Resultsmentioning

confidence: 98%

“…Minimally glycosylated N‐ and C‐domain human ACE proteins (N389 and G13 respectively) were generated by expression in cultured mammalian CHO cells and purified to homogeneity as described previously 20, 31. Synthesis and inhibition data for sampatrilat and samAsp analogue have been reported previously 18.…”

Section: Experimental Methodsmentioning

confidence: 99%

“…Raw data images were indexed and integrated with DIALS 32, and then scaled using AIMLESS 33 from the CCP4 suite 34. Initial phases were obtained by molecular replacement with PHASER 35 using PDB code 1O8A 19 for cACE and 3NXQ 20 for nACE as the search models. Further refinement was initially carried out using REFMAC5 36 and then Phenix 37, with COOT 38 used for rounds of manual model building.…”

Section: Experimental Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Crystal structures of sampatrilat and sampatrilat‐Asp in complex with human ACE – a molecular basis for domain selectivity

et al. 2018

Self Cite

View full text Add to dashboard Cite

Angiotensin‐1‐converting enzyme (ACE) is a zinc metallopeptidase that consists of two homologous catalytic domains (known as nACE and cACE) with different substrate specificities. Based on kinetic studies it was previously reported that sampatrilat, a tight‐binding inhibitor of ACE, K i = 13.8 nm and 171.9 nm for cACE and nACE respectively [Sharma et al., Journal of Chemical Information and Modeling (2016), 56, 2486–2494], was 12.4‐fold more selective for cACE. In addition, samAsp, in which an aspartate group replaces the sampatrilat lysine, was found to be a nonspecific and lower micromolar affinity inhibitor. Here, we report a detailed three‐dimensional structural analysis of sampatrilat and samAsp binding to ACE using high‐resolution crystal structures elucidated by X‐ray crystallography, which provides a molecular basis for differences in inhibitor affinity and selectivity for nACE and cACE. The structures show that the specificity of sampatrilat can be explained by increased hydrophobic interactions and a H‐bond from Glu403 of cACE with the lysine side chain of sampatrilat that are not observed in nACE. In addition, the structures clearly show a significantly greater number of hydrophilic and hydrophobic interactions with sampatrilat compared to samAsp in both cACE and nACE consistent with the difference in affinities. Our findings provide new experimental insights into ligand binding at the active site pockets that are important for the design of highly specific domain selective inhibitors of ACE.DatabaseThe atomic coordinates and structure factors for N‐ and C‐domains of ACE bound to sampatrilat and sampatrilat‐Asp complexes (6F9V, 6F9R, 6F9T and 6F9U respectively) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

show abstract

“…Our extensive knowledge from high resolution structural studies on how inhibitors interact with both the N- and C-domains of human ACE and insect ACE2549505152535455565758 has revealed the molecular basis of binding features of the inhibitors and highlights differences in the inhibitor-enzyme interaction of insect ACE compared with the active sites of the human enzyme. Using the AnoACE homology models and amino acid sequence alignment, it is possible to compare different ACE proteins to look for residue and environment differences in both AnoACE2 and AnoACE3, which could be targeted to create specific inhibitors against these enzymes.…”

Section: Resultsmentioning

confidence: 99%

The toxicity of angiotensin converting enzyme inhibitors to larvae of the disease vectors Aedes aegypti and Anopheles gambiae

Hasan

Williams

Ismail

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

The control of mosquitoes is threatened by the appearance of insecticide resistance and therefore new control chemicals are urgently required. Here we show that inhibitors of mosquito peptidyl dipeptidase, a peptidase related to mammalian angiotensin-converting enzyme (ACE), are insecticidal to larvae of the mosquitoes, Aedes aegypti and Anopheles gambiae. ACE inhibitors (captopril, fosinopril and fosinoprilat) and two peptides (trypsin-modulating oostatic factor/TMOF and a bradykinin-potentiating peptide, BPP-12b) were all inhibitors of the larval ACE activity of both mosquitoes. Two inhibitors, captopril and fosinopril (a pro-drug ester of fosinoprilat), were tested for larvicidal activity. Within 24 h captopril had killed >90% of the early instars of both species with 3rd instars showing greater resistance. Mortality was also high within 24 h of exposure of 1st, 2nd and 3rd instars of An. gambiae to fosinopril. Fosinopril was also toxic to Ae. aegypti larvae, although the 1st instars appeared to be less susceptible to this pro-drug even after 72 h exposure. Homology models of the larval An. gambiae ACE proteins (AnoACE2 and AnoACE3) reveal structural differences compared to human ACE, suggesting that structure-based drug design offers a fruitful approach to the development of selective inhibitors of mosquito ACE enzymes as novel larvicides.

show abstract

The N Domain of Human Angiotensin-I-converting Enzyme

Cited by 77 publications

References 37 publications

Fragment-based design for the development of N-domain-selective angiotensin-1-converting enzyme inhibitors

Fragment-based design for the development of N-domain-selective angiotensin-1-converting enzyme inhibitors

Crystal structures of sampatrilat and sampatrilat‐Asp in complex with human ACE – a molecular basis for domain selectivity

The toxicity of angiotensin converting enzyme inhibitors to larvae of the disease vectors Aedes aegypti and Anopheles gambiae

Contact Info

Product

Resources

About