{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:35:42Z","timestamp":1787027742010,"version":"build-2736575974"},"reference-count":72,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,12]],"date-time":"2024-08-12T00:00:00Z","timestamp":1723420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"CARIPARO Foundation Excellence Research Project","award":["BPiTA n. 52012"],"award-info":[{"award-number":["BPiTA n. 52012"]}]},{"name":"CARIPARO Foundation Excellence Research Project","award":["PRIN-2022 n. 2022ZSA2JP"],"award-info":[{"award-number":["PRIN-2022 n. 2022ZSA2JP"]}]},{"name":"CARIPARO Foundation Excellence Research Project","award":["PRIN-2022 n. 2022FPWYA8"],"award-info":[{"award-number":["PRIN-2022 n. 2022FPWYA8"]}]},{"name":"Italian Ministry of University","award":["BPiTA n. 52012"],"award-info":[{"award-number":["BPiTA n. 52012"]}]},{"name":"Italian Ministry of University","award":["PRIN-2022 n. 2022ZSA2JP"],"award-info":[{"award-number":["PRIN-2022 n. 2022ZSA2JP"]}]},{"name":"Italian Ministry of University","award":["PRIN-2022 n. 2022FPWYA8"],"award-info":[{"award-number":["PRIN-2022 n. 2022FPWYA8"]}]},{"name":"Research (MIUR)","award":["BPiTA n. 52012"],"award-info":[{"award-number":["BPiTA n. 52012"]}]},{"name":"Research (MIUR)","award":["PRIN-2022 n. 2022ZSA2JP"],"award-info":[{"award-number":["PRIN-2022 n. 2022ZSA2JP"]}]},{"name":"Research (MIUR)","award":["PRIN-2022 n. 2022FPWYA8"],"award-info":[{"award-number":["PRIN-2022 n. 2022FPWYA8"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomimetics"],"abstract":"<jats:p>Tick Anticoagulant Peptide (TAP), a 60-amino acid protein from the soft tick Ornithodoros moubata, inhibits activated coagulation factor X (fXa) with almost absolute specificity. Despite TAP and Bovine Pancreatic Trypsin Inhibitor (BPTI) (i.e., the prototype of the Kunitz-type protease inhibitors) sharing a similar 3D fold and disulphide bond topology, they have remarkably different amino acid sequence (only ~24% sequence identity), thermal stability, folding pathways, protease specificity, and even mechanism of protease inhibition. Here, fully active and correctly folded TAP was produced in reasonably high yields (~20%) by solid-phase peptide chemical synthesis and thoroughly characterised with respect to its chemical identity, disulphide pairing, folding kinetics, conformational dynamics, and fXa inhibition. The versatility of the chemical synthesis was exploited to perform structure\u2013activity relationship studies on TAP by incorporating non-coded amino acids at positions 1 and 3 of the inhibitor. Using Hydrogen\u2013Deuterium Exchange Mass Spectrometry, we found that TAP has a remarkably higher conformational flexibility compared to BPTI, and propose that these different dynamics could impact the different folding pathway and inhibition mechanisms of TAP and BPTI. Hence, the TAP\/BPTI pair represents a nice example of divergent evolution, while the relative facility of TAP synthesis could represent a good starting point to design novel synthetic analogues with improved pharmacological profiles.<\/jats:p>","DOI":"10.3390\/biomimetics9080485","type":"journal-article","created":{"date-parts":[[2024,8,12]],"date-time":"2024-08-12T11:23:46Z","timestamp":1723461826000},"page":"485","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Chemical Synthesis and Structure\u2013Activity Relationship Studies of the Coagulation Factor Xa Inhibitor Tick Anticoagulant Peptide from the Hematophagous Parasite Ornithodoros moubata"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9775-8894","authenticated-orcid":false,"given":"Vincenzo","family":"De Filippis","sequence":"first","affiliation":[{"name":"Laboratory of Protein Chemistry & Molecular Haematology, Department of Pharmaceutical and Pharmacological Sciences, School of Medicine, University of Padova, Via F. Marzolo 5, 35131 Padua, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6495-0871","authenticated-orcid":false,"given":"Laura","family":"Acquasaliente","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Chemistry & Molecular Haematology, Department of Pharmaceutical and Pharmacological Sciences, School of Medicine, University of Padova, Via F. Marzolo 5, 35131 Padua, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9396-619X","authenticated-orcid":false,"given":"Andrea","family":"Pierangelini","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Chemistry & Molecular Haematology, Department of Pharmaceutical and Pharmacological Sciences, School of Medicine, University of Padova, Via F. Marzolo 5, 35131 Padua, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6175-4039","authenticated-orcid":false,"given":"Oriano","family":"Marin","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, School of Medicine, University of Padova, Via Trieste 75, 35121 Padua, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1160\/TH09-04-0221","article-title":"Molecular diversity of anticoagulants from haematophagous animals","volume":"102","author":"Koh","year":"2009","journal-title":"Thromb. Haemost."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/j.ibmb.2009.07.003","article-title":"Tick-derived Kunitz-type inhibitors as antihemostatic factors","volume":"39","year":"2009","journal-title":"Insect Biochem. Mol. 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