{"id":6666,"date":"2023-02-17T07:07:34","date_gmt":"2023-02-17T07:07:34","guid":{"rendered":"https:\/\/beta.nanion.de\/resource_library\/2022-the-human-trpa1-intrinsic-cold-and-heat-sensitivity-involves-separate-channel-structures-beyond-the-n-ard-domain\/"},"modified":"2023-02-17T07:07:34","modified_gmt":"2023-02-17T07:07:34","slug":"2022-the-human-trpa1-intrinsic-cold-and-heat-sensitivity-involves-separate-channel-structures-beyond-the-n-ard-domain","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/zh-hans\/resource_library\/2022-the-human-trpa1-intrinsic-cold-and-heat-sensitivity-involves-separate-channel-structures-beyond-the-n-ard-domain\/","title":{"rendered":"2022 &#8211; The human TRPA1 intrinsic cold and heat sensitivity involves separate channel structures beyond the N-ARD domain"},"content":{"rendered":"<p>The human TRPA1 (hTRPA1) is an intrinsic thermosensitive ion channel responding to both\u00a0cold and heat, depending on the redox environment. Here, we have studied purified hTRPA1\u00a0truncated proteins to gain further insight into the temperature gating of hTRPA1. We found in\u00a0patch-clamp bilayer recordings that \u22061-688 hTRPA1, without the N-terminal ankyrin repeat\u00a0domain (N-ARD), was more sensitive to cold and heat, whereas \u22061-854 hTRPA1 that is also\u00a0lacking the S1-S4 voltage sensing-like domain (VSLD) gained sensitivity to cold but lost its\u00a0heat sensitivity. The thiol reducing agent TCEP abolished the temperature sensitivity of both\u00a0\u22061-688 hTRPA1 and \u22061-854 hTRPA1. Cold and heat activity of \u22061-688 hTRPA1 and \u22061-854\u00a0hTRPA1 were associated with different structural conformational changes as revealed by\u00a0intrinsic tryptophan fluorescence measurements. Heat evoked major structural rearrangement\u00a0of the VSLD as well as the C-terminus domain distal to the transmembrane pore domain S5-S6\u00a0(CTD), whereas cold only caused minor conformational changes. As shown for \u03941-854\u00a0hTRPA1, a sudden drop in tryptophan fluorescence occurred within 25-20\u00b0C indicating a\u00a0transition between heat and cold conformations of the CTD, and thus it is proposed that the\u00a0CTD contains a bidirectional temperature switch priming hTRPA1 for either cold or heat. In\u00a0whole-cell patch clamp electrophysiology experiments, replacement of the cysteines 865, 1021\u00a0and 1025 with alanine modulated the cold sensitivity of hTRPA1 when heterologously\u00a0expressed in HEK293T cells. It is proposed that the hTRPA1 CTD harbors cold and heat\u00a0sensitive domains allosterically coupled to the S5-S6 pore region and the VSLD, respectively.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[45,129],"resource_type":[75],"application_areas":[50],"therapeutic_areas":[],"target_categories":[337,338],"target_families":[339],"targets":[126],"resource_keyword":[],"class_list":["post-6666","resource_library","type-resource_library","status-publish","hentry","products_tags-port-a-patch","products_tags-vesicle-prep-pro","resource_type-publication","application_areas-ion-channels","target_categories-ligand-gated-ion-channels","target_categories-other-ion-channels","target_families-trp-channels","targets-trpa1"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library\/6666","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library"}],"about":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/types\/resource_library"}],"wp:attachment":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/media?parent=6666"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/products_tags?post=6666"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_type?post=6666"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/application_areas?post=6666"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/therapeutic_areas?post=6666"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_categories?post=6666"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_families?post=6666"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/targets?post=6666"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_keyword?post=6666"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}