{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T01:13:43Z","timestamp":1790817223535,"version":"4.1.0"},"reference-count":73,"publisher":"SAGE Publications","issue":"1-3","license":[{"start":{"date-parts":[[2022,1,24]],"date-time":"2022-01-24T00:00:00Z","timestamp":1642982400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2022,1,24]],"date-time":"2022-01-24T00:00:00Z","timestamp":1642982400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Antioxidants &amp; Redox Signaling"],"published-print":{"date-parts":[[2022,7]]},"abstract":"<jats:sec>\n                    <jats:title>Aims:<\/jats:title>\n                    <jats:p>\n                      Genetically encoded green fluorescent protein (GFP)-based redox biosensors are widely used to monitor specific and dynamic redox processes in living cells. Over the last few years, various biosensors for a variety of applications were engineered and enhanced to match the organism and cellular environments, which should be investigated. In this context, the unicellular intraerythrocytic parasite\n                      <jats:italic toggle=\"yes\">Plasmodium<\/jats:italic>\n                      , the causative agent of malaria, represents a challenge, as the small size of the organism results in weak fluorescence signals that complicate precise measurements, especially for cell compartment-specific observations. To address this, we have functionally and structurally characterized an enhanced redox biosensor superfolder roGFP2 (sfroGFP2).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      SfroGFP2 retains roGFP2-like behavior, yet with improved fluorescence intensity (FI)\n                      <jats:italic toggle=\"yes\">in cellulo<\/jats:italic>\n                      . SfroGFP2-based redox biosensors are pH insensitive in a physiological pH range and show midpoint potentials comparable with roGFP2-based redox biosensors. Using crystallography and rigidity theory, we identified the superfolding mutations as being responsible for improved structural stability of the biosensor in a redox-sensitive environment, thus explaining the improved FI\n                      <jats:italic toggle=\"yes\">in cellulo<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Innovation:<\/jats:title>\n                    <jats:p>This work provides insight into the structure and function of GFP-based redox biosensors. It describes an improved redox biosensor (sfroGFP2) suitable for measuring oxidizing effects within small cells where applicability of other redox sensor variants is limited.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion:<\/jats:title>\n                    <jats:p>\n                      Improved structural stability of sfroGFP2 gives rise to increased FI\n                      <jats:italic toggle=\"yes\">in cellulo<\/jats:italic>\n                      . Fusion to hGrx1 (human glutaredoxin-1) provides the hitherto most suitable biosensor for measuring oxidizing effects in\n                      <jats:italic toggle=\"yes\">Plasmodium<\/jats:italic>\n                      . This sensor is of major interest for studying glutathione redox changes in small cells, as well as subcellular compartments in general.\n                      <jats:italic toggle=\"yes\">Antioxid. Redox Signal.<\/jats:italic>\n                      37, 1\u201318.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1089\/ars.2021.0234","type":"journal-article","created":{"date-parts":[[2022,1,24]],"date-time":"2022-01-24T10:40:15Z","timestamp":1643020815000},"page":"1-18","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":16,"title":["Structure and Function of Redox-Sensitive Superfolder Green Fluorescent Protein Variant"],"prefix":"10.1177","volume":"37","author":[{"given":"Kim C.","family":"Heimsch","sequence":"first","affiliation":[{"name":"Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoph G.W.","family":"Gertzen","sequence":"additional","affiliation":[{"name":"Institute for Pharmaceutical and Medicinal Chemistry, Heinrich Heine University D\u00fcsseldorf, D\u00fcsseldorf, Germany."},{"name":"Center for Structural Studies (CSS), Heinrich Heine University D\u00fcsseldorf, D\u00fcsseldorf, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna Katharina","family":"Schuh","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Nietzel","sequence":"additional","affiliation":[{"name":"Institute of Plant Biology and Biotechnology, University of M\u00fcnster, M\u00fcnster, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Rahlfs","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jude M.","family":"Przyborski","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holger","family":"Gohlke","sequence":"additional","affiliation":[{"name":"Institute for Pharmaceutical and Medicinal Chemistry, Heinrich Heine University D\u00fcsseldorf, D\u00fcsseldorf, Germany."},{"name":"John von Neumann Institute of Computing (NIC), J\u00fclich Supercomputing Centre (JSC), Institute of Biological Information Processing (IBI-7: Structural Biochemistry), and Institute of Bio- and Geosciences (IBG-4: Bioinformatics), Forschungszentrum J\u00fclich GmbH, J\u00fclich, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus","family":"Schwarzl\u00e4nder","sequence":"additional","affiliation":[{"name":"Institute of Plant Biology and Biotechnology, University of M\u00fcnster, M\u00fcnster, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katja","family":"Becker","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4422-6795","authenticated-orcid":false,"given":"Karin","family":"Fritz-Wolf","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen, Germany."},{"name":"Max-Planck Institute of Medical Research, Heidelberg, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2022,1,24]]},"reference":[{"key":"e_1_3_6_2_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"e_1_3_6_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.22102"},{"key":"e_1_3_6_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.6.2306"},{"key":"e_1_3_6_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2017.12.006"},{"key":"e_1_3_6_6_2","volume-title":"AMBER 2018","author":"Case DA","year":"2018","unstructured":"Case DA, , Brozell SR, , Cerutti DS, , Cheatham TE, , Cruzeiro VWD III, , Darden TA, , Duke RE, , Ghoreishi D, , Gohlke H, , Goetz AW, , Greene D, , Harris R, , Homeyer N, , Izadi S, , Kovalenko A, , Lee TS, , LeGrand S, , Li P, , Lin C, , Liu J, , Luchko T, , Luo R, , Mermelstein DJ, , Merz KM, , Miao Y, , Monard G, , Nguyen H, , Omelyan I, , Onufriev A, , Pan F, , Qi R, , Roe DR, , Roitberg A, , Sagui C, , Schott-Verdugo S, , Shen J, , Simmerling CL, , Smith J, , Swails J, , Walker RC, , Wang J, , Wei H, , Wolf RM, , Wu X, , Xiao L, , York DM, , and Kollman PA. 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