{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T15:10:52Z","timestamp":1788016252436,"version":"build-2784847793"},"reference-count":40,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2023,6,16]],"date-time":"2023-06-16T00:00:00Z","timestamp":1686873600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2023,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>As a consequence of the fast resistance spreading, a limited number of drugs are available to treat fungal infections. Therefore, there is an urgent need to develop new antifungal treatment strategies. The features of a disulfide bond\u2010stabilized antifungal protein, NFAP2 secreted by the mold <jats:italic>Neosartorya<\/jats:italic> (<jats:italic>Aspergillus<\/jats:italic>) <jats:italic>fischeri<\/jats:italic> render it to be a promising template for future protein\u2010based antifungal drug design, which requires knowledge about the native disulfide linkage pattern as it is one of the prerequisites for biological activity. However, in the lack of tryptic and chymotryptic proteolytic sites in the ACNCPNNCK sequence, the determination of the disulfide linkage pattern of NFAP2 is not easy with traditional mass spectrometry\u2010based methods. According to <jats:italic>in silico<\/jats:italic> predictions working with a preliminary nuclear magnetic resonance (NMR) solution structure, two disulfide isomers of NFAP2 (<jats:italic>abbacc<\/jats:italic> and <jats:italic>abbcac<\/jats:italic>) were possible. Both were chemically synthesized; and comparative reversed\u2010phase high\u2010performance liquid chromatography, electronic circular dichroism and NMR spectroscopy analyses, and antifungal susceptibility and efficacy tests indicated that the <jats:italic>abbcac<\/jats:italic> is the native pattern. This knowledge allowed rational modification of NAFP2 to improve the antifungal efficacy and spectrum through the modulation of the evolutionarily conserved \u03b3\u2010core region, which is responsible for the activity of several antimicrobial peptides. Disruption of the steric structure of NFAP2 upon \u03b3\u2010core modification led to the conclusions that this motif may affect the formation of the biologically active three\u2010dimensional structure, and that the \u03b3\u2010core modulation is not an efficient tool to improve the antifungal efficacy or to change the antifungal spectrum of NFAP2.<\/jats:p>","DOI":"10.1002\/pro.4692","type":"journal-article","created":{"date-parts":[[2023,6,7]],"date-time":"2023-06-07T02:49:06Z","timestamp":1686106146000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Hard nut to crack: Solving the disulfide linkage pattern of the <i>Neosartorya<\/i> (<i>Aspergillus<\/i>) <i>fischeri<\/i> antifungal protein 2"],"prefix":"10.1002","volume":"32","author":[{"given":"Gy\u00f6rgyi","family":"V\u00e1radi","sequence":"first","affiliation":[{"name":"Department of Medical Chemistry Albert Szent\u2010Gy\u00f6rgyi Medical School, University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zolt\u00e1n","family":"Kele","sequence":"additional","affiliation":[{"name":"Department of Medical Chemistry Albert Szent\u2010Gy\u00f6rgyi Medical School, University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e1s","family":"Czajlik","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Science and Technology University of Debrecen  Debrecen Hungary"},{"name":"Department of Biochemistry Institute of Biochemistry and Molecular Biology, Semmelweis University  Budapest Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Attila","family":"Borics","sequence":"additional","affiliation":[{"name":"Laboratory of Chemical Biology Institute of Biochemistry, Biological Research Centre, E\u00f6tv\u00f6s Lor\u00e1nd Research Network  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e1bor","family":"Bende","sequence":"additional","affiliation":[{"name":"Department of Biotechnology Faculty of Science and Informatics, University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Csaba","family":"Papp","sequence":"additional","affiliation":[{"name":"Department of Microbiology Faculty of Science and Informatics, University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e1bor","family":"R\u00e1khely","sequence":"additional","affiliation":[{"name":"Department of Biotechnology Faculty of Science and Informatics, University of Szeged  Szeged Hungary"},{"name":"Institute of Biophysics, Biological Research Centre, E\u00f6tv\u00f6s Lor\u00e1nd Research Network  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e1bor K.","family":"T\u00f3th","sequence":"additional","affiliation":[{"name":"Department of Medical Chemistry Albert Szent\u2010Gy\u00f6rgyi Medical School, University of Szeged  Szeged Hungary"},{"name":"MTA\u2010SZTE Biomimetic Systems Research Group, University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gyula","family":"Batta","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Science and Technology University of Debrecen  Debrecen Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6976-8910","authenticated-orcid":false,"given":"L\u00e1szl\u00f3","family":"Galg\u00f3czy","sequence":"additional","affiliation":[{"name":"Department of Biotechnology Faculty of Science and Informatics, University of Szeged  Szeged Hungary"},{"name":"Fungal Genomics and Evolution Lab, Institute of Biochemistry, Biological Research Centre, E\u00f6tv\u00f6s Lor\u00e1nd Research Network  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,6,16]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2009.07011.x"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkl266"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-14-346"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22031183"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.2147\/IJN.S187957"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.00105"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkl189"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201404879"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0204825"},{"key":"e_1_2_7_11_1","unstructured":"Fortune Business Insights.Report ID: FBI101188. 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