{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T14:10:52Z","timestamp":1760278252884,"version":"build-2065373602"},"reference-count":52,"publisher":"Wiley","issue":"28","license":[{"start":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T00:00:00Z","timestamp":1745193600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001502","name":"Department of Atomic Energy, Government of India","doi-asserted-by":"publisher","award":["RTI\u20104007"],"award-info":[{"award-number":["RTI\u20104007"]}],"id":[{"id":"10.13039\/501100001502","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Chemistry A European J"],"published-print":{"date-parts":[[2025,5,19]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The interaction between apical membrane antigen 1 (<jats:italic>Pf<\/jats:italic>AMA1) and rhoptry neck protein 2 (<jats:italic>Pf<\/jats:italic>RON2) is crucial for <jats:italic>Plasmodium falciparum<\/jats:italic> red blood cell invasion, making it a key target for anti\u2010malarial drug development strategies. Here, we report the chemical synthesis of <jats:italic>Pf<\/jats:italic>AMA1 domain I (<jats:italic>Pf<\/jats:italic>AMA1\u2010DI) in both linear and backbone\u2010circularized forms, employing a six\u2010segment convergent synthesis approach exploiting one\u2010pot chemistries and solubilizing tags. The chemically synthesized linear <jats:italic>Pf<\/jats:italic>AMA1\u2010DI construct exhibited incomplete disulfide bond formation during folding, likely due to increased terminal flexibility in the absence of domain II. To address this, we employed backbone cyclization of the large 180\u2010residue polypeptide chain, with 3\u2010residue linker sequence, as a unique strategy to conformationally restrict its termini and facilitate correct disulfide bond formation. Introducing a multipurpose affinity and solubility tag to the cyclic<jats:italic>Pf<\/jats:italic>AMA1\u2010DI construct further improved the folding yield by mitigating aggregation. The predicted structure using ColabFold\u2010Alphafold2 indicated that <jats:italic>Pf<\/jats:italic>RON2 ligand binds within the hydrophobic groove of the cyclic<jats:italic>Pf<\/jats:italic>AMA1\u2010DI construct similar to the native interactions. These findings underscore the potential of large protein backbone cyclization to stabilize protein structure, offering a compelling strategy for the chemical synthesis of otherwise unstable protein domains with broad applications in miniature protein engineering.<\/jats:p>","DOI":"10.1002\/chem.202500894","type":"journal-article","created":{"date-parts":[[2025,4,8]],"date-time":"2025-04-08T11:57:07Z","timestamp":1744113427000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Chemical Protein Engineering: Backbone Cyclization Rescues Folding of a 183\u2010Residue Truncated Domain of Malaria Parasite Protein <i>Pf<\/i>AMA1"],"prefix":"10.1002","volume":"31","author":[{"given":"Jamsad","family":"Mannuthodikayil","sequence":"first","affiliation":[{"name":"Tata Institute of Fundamental Research Hyderabad  36\/p Gopanpally Hyderabad Telangana 500046 India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vishal","family":"Malik","sequence":"additional","affiliation":[{"name":"Tata Institute of Fundamental Research Hyderabad  36\/p Gopanpally Hyderabad Telangana 500046 India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abhisek","family":"Kar","sequence":"additional","affiliation":[{"name":"Tata Institute of Fundamental Research Hyderabad  36\/p Gopanpally Hyderabad Telangana 500046 India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sameer","family":"Singh","sequence":"additional","affiliation":[{"name":"Tata Institute of Fundamental Research Hyderabad  36\/p Gopanpally Hyderabad Telangana 500046 India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3194-1378","authenticated-orcid":false,"given":"Kalyaneswar","family":"Mandal","sequence":"additional","affiliation":[{"name":"Tata Institute of Fundamental Research Hyderabad  36\/p Gopanpally Hyderabad Telangana 500046 India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,4,21]]},"reference":[{"volume-title":"WHO World Malaria Report","year":"2024","author":"Global Malaria Programme (GMP) Team","key":"e_1_2_9_1_1"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001276"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0109674"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms3261"},{"key":"e_1_2_9_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10822-015-9842-7"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M207985200"},{"key":"e_1_2_9_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.10.6981-6989.2005"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2016.07.001"},{"key":"e_1_2_9_6_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202200533"},{"key":"e_1_2_9_6_3","unstructured":"c)A.Biswas A.Narayan S.Sinha K.Mandal bioRxiv preprint 2023 2023.2006.2023.546305;"},{"key":"e_1_2_9_6_4","doi-asserted-by":"publisher","DOI":"10.1039\/D4CB00229F"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002755"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0144764"},{"key":"e_1_2_9_9_1","volume":"26","author":"Biswas A.","year":"2021","journal-title":"Biochem. Biophys. Rep."},{"key":"e_1_2_9_9_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0126206"},{"key":"e_1_2_9_9_3","doi-asserted-by":"publisher","DOI":"10.1186\/s12936-019-2649-6"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.271.5257.1854"},{"key":"e_1_2_9_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200300570"},{"key":"e_1_2_9_10_3","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1210483109"},{"key":"e_1_2_9_10_4","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.4c12057"},{"key":"e_1_2_9_10_5","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202414256"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41570-023-00493-y"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1039\/c39780000537"},{"key":"e_1_2_9_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja00897a025"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.7973629"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201600101"},{"key":"e_1_2_9_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200353540"},{"key":"e_1_2_9_14_3","doi-asserted-by":"publisher","DOI":"10.1016\/S0040-4039(00)89595-7"},{"key":"e_1_2_9_14_4","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202000491"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200704195"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0282(1999)51:4<247::AID-BIP2>3.0.CO;2-W"},{"key":"e_1_2_9_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2013.152"},{"key":"e_1_2_9_16_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201100996"},{"key":"e_1_2_9_16_4","doi-asserted-by":"publisher","DOI":"10.1021\/acs.orglett.9b03440"},{"key":"e_1_2_9_16_5","doi-asserted-by":"publisher","DOI":"10.1007\/s11426-016-0381-1"},{"key":"e_1_2_9_16_6","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200705471"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/9783527823567.ch7"},{"key":"e_1_2_9_17_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7SC00488E"},{"key":"e_1_2_9_17_3","doi-asserted-by":"publisher","DOI":"10.1016\/j.tetlet.2007.01.030"},{"key":"e_1_2_9_17_4","doi-asserted-by":"publisher","DOI":"10.1002\/psc.634"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abb2491"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0501808102"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms241713257"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.6b04310"},{"key":"e_1_2_9_21_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7SC00789B"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2003.09.008"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(82)90515-0"},{"key":"e_1_2_9_24_1","doi-asserted-by":"crossref","unstructured":"J. M.Collins S. K.Singh US Patent App. 14\/969 004 2016.","DOI":"10.1145\/2967510"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202000865"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201106060"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1039\/D3CB00208J"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202400894"}],"container-title":["Chemistry \u2013 A European Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/chem.202500894","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T13:28:49Z","timestamp":1760275729000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/chem.202500894"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,21]]},"references-count":52,"journal-issue":{"issue":"28","published-print":{"date-parts":[[2025,5,19]]}},"alternative-id":["10.1002\/chem.202500894"],"URL":"https:\/\/doi.org\/10.1002\/chem.202500894","archive":["Portico"],"relation":{},"ISSN":["0947-6539","1521-3765"],"issn-type":[{"type":"print","value":"0947-6539"},{"type":"electronic","value":"1521-3765"}],"subject":[],"published":{"date-parts":[[2025,4,21]]},"assertion":[{"value":"2025-03-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202500894"}}