{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T16:01:49Z","timestamp":1790784109803,"version":"4.1.0"},"reference-count":93,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,10,19]],"date-time":"2020-10-19T00:00:00Z","timestamp":1603065600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR-10-LABX-12-01, ANR-17-CE11-0022-01, ANR-10-INBS-04-01, ANR-10-INBS-05"],"award-info":[{"award-number":["ANR-10-LABX-12-01, ANR-17-CE11-0022-01, ANR-10-INBS-04-01, ANR-10-INBS-05"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["648030"],"award-info":[{"award-number":["648030"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>The high-resolution structural study of huntingtin exon-1 (HttEx1) has long been hampered by its intrinsic properties. In addition to being prone to aggregate, HttEx1 contains low-complexity regions (LCRs) and is intrinsically disordered, ruling out several standard structural biology approaches. Here, we use a cell-free (CF) protein expression system to robustly and rapidly synthesize (sub-) pathological HttEx1. The open nature of the CF reaction allows the application of different isotopic labeling schemes, making HttEx1 amenable for nuclear magnetic resonance studies. While uniform and selective labeling facilitate the sequential assignment of HttEx1, combining CF expression with nonsense suppression allows the site-specific incorporation of a single labeled residue, making possible the detailed investigation of the LCRs. To optimize CF suppression yields, we analyze the expression and suppression kinetics, revealing that high concentrations of loaded suppressor tRNA have a negative impact on the final reaction yield. The optimized CF protein expression and suppression system is very versatile and well suited to produce challenging proteins with LCRs in order to enable the characterization of their structure and dynamics.<\/jats:p>","DOI":"10.3390\/biom10101458","type":"journal-article","created":{"date-parts":[[2020,10,19]],"date-time":"2020-10-19T09:36:28Z","timestamp":1603100188000},"page":"1458","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Robust Cell-Free Expression of Sub-Pathological and Pathological Huntingtin Exon-1 for NMR Studies. General Approaches for the Isotopic Labeling of Low-Complexity Proteins"],"prefix":"10.3390","volume":"10","author":[{"given":"Anna","family":"Morat\u00f3","sequence":"first","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8244-5676","authenticated-orcid":false,"given":"Carlos A.","family":"Elena-Real","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matija","family":"Popovic","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aur\u00e9lie","family":"Fournet","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen","family":"Zhang","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fr\u00e9d\u00e9ric","family":"Allemand","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8145-6795","authenticated-orcid":false,"given":"Nathalie","family":"Sibille","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annika","family":"Urbanek","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pau","family":"Bernad\u00f3","sequence":"additional","affiliation":[{"name":"Centre de Biochimie Structurale (CBS), INSERM, CNRS and Universit\u00e9 de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/S0140-6736(07)60111-1","article-title":"Huntington\u2019s disease","volume":"369","author":"Walker","year":"2007","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1152\/physrev.00041.2009","article-title":"Molecular mechanisms and potential therapeutical targets in Huntington\u2019s disease","volume":"90","author":"Zuccato","year":"2010","journal-title":"Physiol. 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