{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:10:24Z","timestamp":1760058624105,"version":"build-2065373602"},"reference-count":38,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,4,16]],"date-time":"2025-04-16T00:00:00Z","timestamp":1744761600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"UKRI-Biotechnology and Biological Sciences Research Council Research","award":["BB\/X001032\/1"],"award-info":[{"award-number":["BB\/X001032\/1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Human angiotensin-I-converting enzyme (ACE) is involved in vasoregulation, inflammation, and neurodegenerative disorders. The enzyme is formed of two domains; the C-domain (cACE) is primarily involved in blood pressure regulation, whereas the N-domain (nACE) is strongly linked to fibrosis; hence, designing domain-specific inhibitors could make a difference between treating one condition without having a negative effect on another. AnCE (a close homologue of ACE) is derived from Drosophila melanogaster and has a high similarity specifically to cACE. Due to high similarity and ease of crystallisation, AnCE has been chosen as a model protein for ACE studies and for the design of ACE inhibitors. In this study, enzyme kinetic assays and X-ray crystallography techniques revealed the significance of using dipeptides as selective inhibitors for AnCE and how this knowledge could be applied to cACE and nACE. All the dipeptides tested in this study were shown to bind AnCE in two distinct locations, i.e., the non-prime and prime subsites. It was found that a hydrophobic residue at the S1 and S1\u2032 subsites, with a tryptophan at the S2 and S2\u2032 subsites, showed highest affinity towards AnCE. It was also observed that a key pocket within the S2\u2032 subsite had a major influence on the binding orientation within the prime subsites and could potentially explain ACE\u2019s dipeptidyl carboxypeptidase activity. Importantly these dipeptides are found in functional foods, making them potentially available from diets. Knowledge of the dipeptide binding presented here could aid in the development of ACE domain-specific inhibitors.<\/jats:p>","DOI":"10.3390\/biom15040591","type":"journal-article","created":{"date-parts":[[2025,4,16]],"date-time":"2025-04-16T06:12:06Z","timestamp":1744783926000},"page":"591","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Molecular Basis of Dipeptide Recognition in Drosophila melanogaster Angiotensin I-Converting Enzyme Homologue, AnCE"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-3668-7211","authenticated-orcid":false,"given":"Joanna","family":"\u017bukowska","sequence":"first","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5371-4283","authenticated-orcid":false,"given":"Kyle S.","family":"Gregory","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adam","family":"Robinson","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4792-6559","authenticated-orcid":false,"given":"R. Elwyn","family":"Isaac","sequence":"additional","affiliation":[{"name":"Institute of Integrative and Comparative Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3009-4058","authenticated-orcid":false,"given":"K. Ravi","family":"Acharya","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"5942","DOI":"10.1021\/bi8002605","article-title":"Probing the Basis of Domain-Dependent Inhibition Using Novel Ketone Inhibitors of Angiotensin-Converting Enzyme","volume":"47","author":"Watermeyer","year":"2008","journal-title":"Biochemistry"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"845","DOI":"10.2174\/092986712799034950","article-title":"Structure Based Drug Design of Angiotensin-I Converting Enzyme Inhibitors","volume":"19","author":"Anthony","year":"2012","journal-title":"Curr. Med. 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