{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T14:37:12Z","timestamp":1754145432486,"version":"3.41.2"},"reference-count":54,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2022,8,22]],"date-time":"2022-08-22T00:00:00Z","timestamp":1661126400000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2022,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>This article outlines the design and development of scyllatoxin (ScTx)\u2013based BH3 domain mimetics with diverse patterns of native disulfide bonds. More specifically, this method summarizes the total chemical synthesis of ScTx\u2010based peptides that contain zero, one, two, or three disulfide linkages, including techniques to generate variants with any combination of native disulfides. Each peptide reported herein is generated on solid\u2010phase support using microwave\u2010assisted coupling procedures, and all reaction parameters related to the peptide synthesis are described in detail. The various disulfide patterns of the ScTx\u2010based constructs are established during peptide synthesis and are ultimately verified by mass analysis of trypsin\u2010digested fragments. The BH3 domain mimetics developed herein were generated by transposing residues from the helical BH3 domain of the pro\u2010apoptotic BCL2 protein Bax to the \u03b1\u2010helix of wild\u2010type ScTx. Interestingly, we found that the relative binding affinities of ScTx\u2010Bax peptides for the anti\u2010apoptotic BCL2 protein Bcl\u20102 (proper) were heavily influenced by the number and position of disulfide linkages within the ScTx\u2010Bax sequence. As a consequence, we were able to utilize ScTx\u2010Bax BH3 domain mimetics with varied patterns of disulfide bonds to survey how structural rigidity within the helical Bax BH3 domain affects binding to promiscuous anti\u2010apoptotic BCL2 proteins. More broadly, the ability to generate ScTx\u2010based molecules that contain any combination of native disulfide bonds expands the utility of such constructs as tools to study the molecular nature of protein\u2010protein interactions. \u00a9 2022 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Synthesis and characterization of ScTx\u2010based Bax BH3 domain mimetics<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Oxidation of ScTx\u2010Bax BH3 domain mimetics containing one, two, or three disulfide linkages<\/jats:p><jats:p><jats:bold>Support Protocol<\/jats:bold>: Mapping of disulfide linkages in oxidized ScTx\u2010Bax BH3 domain mimetics<\/jats:p>","DOI":"10.1002\/cpz1.526","type":"journal-article","created":{"date-parts":[[2022,8,22]],"date-time":"2022-08-22T14:04:13Z","timestamp":1661177053000},"source":"Crossref","is-referenced-by-count":1,"title":["Synthesis of Scyllatoxin\u2010Based BH3 Domain Mimetics with Diverse Patterns of Native Disulfide Bonds"],"prefix":"10.1002","volume":"2","author":[{"given":"Matthew J. K.","family":"Vince","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry Ohio University  Athens Ohio"},{"name":"Institut f\u00fcr Bioanalytische Chemie, Fakult\u00e4t f\u00fcr Chemie und Mineralogie Universit\u00e4t Leipzig  Leipzig Germany"},{"name":"Biotechnologisch\u2010Biomedizinisches Zentrum Universit\u00e4t Leipzig  Leipzig Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Justin M.","family":"Holub","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Ohio University  Athens Ohio"},{"name":"Edison Biotechnology Institute Ohio University  Athens Ohio"},{"name":"Molecular and Cellular Biology Program Ohio University  Athens Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2022,8,22]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10930\u2010018\u20109791\u20109"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/psc.2999"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.5b04563"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.16.1.395"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2004.12.030"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi000096"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1207102"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm3722"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1110\/ps.073105408"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13277\u2010015\u20103719\u20101"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399\u20103011.1990.tb00939.x"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja037954k"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1039\/C5OB02080H"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abb2491"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201900717"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2008.08.019"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/ddr.21408"},{"key":"e_1_2_8_19_1","first-page":"956","article-title":"Synthesis of linear and cyclic disulfide heptapeptides of longicalycinin A and evaluation of toxicity on cancerous cells HepG2 and HT\u201029","volume":"17","author":"Houshdar Tehrani M. 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