{"id":13266,"date":"2023-09-14T13:17:10","date_gmt":"2023-09-14T13:17:10","guid":{"rendered":"https:\/\/www.nanion.de\/?post_type=resource_library&#038;p=13266"},"modified":"2024-01-11T11:47:59","modified_gmt":"2024-01-11T11:47:59","slug":"2023-enhancing-the-performance-of-a-mutant-pyrrolysyl-trna-synthetase-to-create-a-highly-versatile-eukaryotic-cell-free-protein-synthesis-tool","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/ja\/resource_library\/2023-enhancing-the-performance-of-a-mutant-pyrrolysyl-trna-synthetase-to-create-a-highly-versatile-eukaryotic-cell-free-protein-synthesis-tool\/","title":{"rendered":"2023 &#8211; Enhancing the performance of a mutant pyrrolysyl-tRNA synthetase to create a highly versatile eukaryotic cell-free protein synthesis tool"},"content":{"rendered":"<p>Modification of proteins with a broad range of chemical functionalities enables the investigation of protein structure and activity by manipulating polypeptides at single amino acid resolution. Indeed, various functional groups including bulky non-canonical amino acids like strained cyclooctenes could be introduced by the unique features of the binding pocket of the double mutant pyrrolysyl-tRNA synthetase (Y306A, Y384F), but the instable nature of the enzyme limits its application in vivo. Here, we constructed a cell-free protein production system, which increased the overall enzyme stability by combining different reaction compartments. Moreover, a co-expression approach in a one-pot reaction allowed straightforward site-specific fluorescent labeling of the functional complex membrane protein cystic fibrosis transmembrane conductance regulator. Our work provides a versatile platform for introducing various non-canonical amino acids into difficult-to-express proteins for structural and fluorescence based investigation of proteins activity.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[98],"resource_type":[75],"application_areas":[99],"therapeutic_areas":[109,143],"target_categories":[],"target_families":[],"targets":[],"resource_keyword":[],"class_list":["post-13266","resource_library","type-resource_library","status-publish","hentry","products_tags-orbit-16-tc","resource_type-publication","application_areas-pores-and-toxins","therapeutic_areas-respiratory-diseases","therapeutic_areas-assay-development-and-validation"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_library\/13266","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_library"}],"about":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/types\/resource_library"}],"wp:attachment":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/media?parent=13266"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/products_tags?post=13266"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_type?post=13266"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/application_areas?post=13266"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/therapeutic_areas?post=13266"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/target_categories?post=13266"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/target_families?post=13266"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/targets?post=13266"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_keyword?post=13266"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}