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In the primitive bacteriophytochromes this includes refolding of a tongue\u2010like hairpin loop close to the chromophore, one strand of an antiparallel \u03b2\u2010sheet being replaced by an \u03b1\u2010helix. However, the strand sequence in the cyanobacterial phytochrome Cph1 is different from that of previously investigated bacteriophytochromes and has a higher \u03b2\u2010sheet propensity. We confirm here the transition experimentally and estimate minimum helix length using dynamic nuclear polarisation (DNP) magic angle spinning NMR. Sample conditions were optimized for protein DNP NMR studies at high field, yielding Boltzmann enhancements \u03f5<jats:sub>B<\/jats:sub> of 19 at an NMR field of 18.801\u2005T. Selective labelling of Trp, Ile, Arg, and Val residues with <jats:sup>13<\/jats:sup>C and <jats:sup>15<\/jats:sup>N enabled filtering for pairs of labelled amino acids by the 3D CANCOCA technique to identify signals of the motif <jats:sub>483<\/jats:sub>Ile\u2010Val\u2010Arg<jats:sub>485<\/jats:sub> (IVR) present in both sheet and helix. Those signals were assigned for the Pfr state of the protein. Based on the chemical shift pattern, we confirm for Cph1 the formation of a helix covering the IVR motif.<\/jats:p>","DOI":"10.1002\/chem.202402454","type":"journal-article","created":{"date-parts":[[2024,11,14]],"date-time":"2024-11-14T16:19:33Z","timestamp":1731601173000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Changes in Secondary Structure Upon Pr to Pfr Transition in Cyanobacterial Phytochrome Cph1 Detected by DNP NMR"],"prefix":"10.1002","volume":"31","author":[{"given":"Lisa","family":"Gerland","sequence":"first","affiliation":[{"name":"NMR-supported Structural Biology Leibniz-Forschungsinstitut f\u00fcr Molekulare Pharmakologie  Robert-R\u00f6ssle-Str. 10 13125 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anne","family":"Diehl","sequence":"additional","affiliation":[{"name":"NMR-supported Structural Biology Leibniz-Forschungsinstitut f\u00fcr Molekulare Pharmakologie  Robert-R\u00f6ssle-Str. 10 13125 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natalja","family":"Erdmann","sequence":"additional","affiliation":[{"name":"NMR-supported Structural Biology Leibniz-Forschungsinstitut f\u00fcr Molekulare Pharmakologie  Robert-R\u00f6ssle-Str. 10 13125 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthias","family":"Hiller","sequence":"additional","affiliation":[{"name":"NMR-supported Structural Biology Leibniz-Forschungsinstitut f\u00fcr Molekulare Pharmakologie  Robert-R\u00f6ssle-Str. 10 13125 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina","family":"Lang","sequence":"additional","affiliation":[{"name":"Plant Physiology Faculty of Biology and Chemistry Justus-Liebig-University Giessen  Senckenbergstr. 3 35390 Giessen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Teutloff","sequence":"additional","affiliation":[{"name":"Department of Physics Freie Universit\u00e4t Berlin  Arnimallee 14 14195 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jon","family":"Hughes","sequence":"additional","affiliation":[{"name":"Plant Physiology Faculty of Biology and Chemistry Justus-Liebig-University Giessen  Senckenbergstr. 3 35390 Giessen Germany"},{"name":"Department of Physics Freie Universit\u00e4t Berlin  Arnimallee 14 14195 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4384-9544","authenticated-orcid":false,"given":"Hartmut","family":"Oschkinat","sequence":"additional","affiliation":[{"name":"NMR-supported Structural Biology Leibniz-Forschungsinstitut f\u00fcr Molekulare Pharmakologie  Robert-R\u00f6ssle-Str. 10 13125 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,11,28]]},"reference":[{"key":"e_1_2_9_1_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-arplant-070623-110636"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-018-0110-4"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1327.2001.02395.x"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/ar800133x"},{"key":"e_1_2_9_5_1","first-page":"1","volume":"2","author":"Escobar F. 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