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In particular, phosphorylation of threonine and serine residues (T3, S13, and\/or S16) has been shown to inhibit Htt aggregation\n                    <jats:italic>in\u2005vitro<\/jats:italic>\n                    and inclusion formation in cellular and animal models of Huntington's disease (HD). In this paper, we describe a new and simple methodology for producing milligram quantities of highly pure wild\u2010type or mutant Httex1 proteins that are site\u2010specifically phosphorylated at T3 or at both S13 and S16. This advance was enabled by 1)\u2005the discovery and validation of novel kinases that efficiently phosphorylate Httex1 at S13 and S16 (TBK1), at T3 (GCK) or T3 and S13 (TNIK and HGK), and 2)\u2005the development of an efficient methodology for producing recombinant native Httex1 proteins by using a SUMO\u2010fusion expression and purification strategy.\n                    <jats:sup>[26]<\/jats:sup>\n                    As a proof of concept, we demonstrate how this method can be applied to produce Httex1 proteins that are both site\u2010specifically phosphorylated and fluorescently or isotopically labeled. Together, these advances should increase access to these valuable tools and expand the range of methods and experimental approaches that can be used to elucidate the mechanisms by which phosphorylation influences Httex1 or HTT structure, aggregation, interactome, and function(s) in health and disease.\n                  <\/jats:p>","DOI":"10.1002\/cbic.202000508","type":"journal-article","created":{"date-parts":[[2020,8,27]],"date-time":"2020-08-27T21:27:27Z","timestamp":1598563647000},"page":"217-231","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Site\u2010Specific Phosphorylation of Huntingtin Exon\u20051 Recombinant Proteins Enabled by the Discovery of Novel Kinases"],"prefix":"10.1002","volume":"22","author":[{"given":"Anass","family":"Chiki","sequence":"first","affiliation":[{"name":"Laboratory of Molecular and Chemical Biology of Neurodegeneration, School of Life Sciences Brain Mind Institute Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Station 19 1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan","family":"Ricci","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular and Chemical Biology of Neurodegeneration, School of Life Sciences Brain Mind Institute Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Station 19 1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ramanath","family":"Hegde","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular and Chemical Biology of Neurodegeneration, School of Life Sciences Brain Mind Institute Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Station 19 1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciano A.","family":"Abriata","sequence":"additional","affiliation":[{"name":"Protein Production and Structure Core Facility and Laboratory for Biomolecular Modeling Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL) and Swiss Institute of Bioinformatics (SIB)  1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas","family":"Reif","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular and Chemical Biology of Neurodegeneration, School of Life Sciences Brain Mind Institute Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Station 19 1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Driss","family":"Boudeffa","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular and Chemical Biology of Neurodegeneration, School of Life Sciences Brain Mind Institute Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Station 19 1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7682-8320","authenticated-orcid":false,"given":"Hilal A.","family":"Lashuel","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular and Chemical Biology of Neurodegeneration, School of Life Sciences Brain Mind Institute Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Station 19 1015 Lausanne Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,10,13]]},"reference":[{"key":"e_1_2_7_1_1","unstructured":"\u00a0"},{"key":"e_1_2_7_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(10)70245-3"},{"key":"e_1_2_7_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/ana.10691"},{"key":"e_1_2_7_4_2","unstructured":"G. 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