{"id":14809,"date":"2024-07-08T10:11:07","date_gmt":"2024-07-08T10:11:07","guid":{"rendered":"https:\/\/www.nanion.de\/?post_type=resource_library&#038;p=14809"},"modified":"2024-07-11T08:12:26","modified_gmt":"2024-07-11T08:12:26","slug":"2024-n-glycosylation-deficiency-in-transgene-%ce%b17-nachr-and-ric3-expressing-cho-cells-without-nacho","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/zh-hans\/resource_library\/2024-n-glycosylation-deficiency-in-transgene-%ce%b17-nachr-and-ric3-expressing-cho-cells-without-nacho\/","title":{"rendered":"2024 &#8211; N-Glycosylation Deficiency in Transgene \u03b17 nAChR and RIC3 Expressing CHO Cells Without NACHO"},"content":{"rendered":"<p>The human neuronal nicotinic acetylcholine receptor \u03b17 (nAChR) is an important target implicated in diseases like Alzheimer\u2019s or Parkinson\u2019s, as well as a validated target for drug discovery. For \u03b17 nAChR model systems, correct folding and ion influx functions are essential. Two chaperones, resistance to inhibitors of cholinesterase 3 (RIC3) and novel nAChR regulator (NACHO), enhance the assembly and function of \u03b17 nAChR. This study investigates the consequence of NACHO absence on \u03b17 nAChR expression and function. Therefore, the sequences of human \u03b17 nAChR and human RIC3 were transduced in Chinese hamster ovary (CHO) cells. Protein expression and function of \u03b17 nAChR were confirmed by Western blot and voltage clamp, respectively. Cellular viability was assessed by cell proliferation and lactate dehydrogenase assays. Intracellular and extracellular expression were determined by in\/on-cell Western, compared with another nAChR subtype by novel cluster fluorescence-linked immunosorbent assay, and N-glycosylation efficiency was assessed by glycosylation digest. The transgene CHO cell line showed expected protein expression and function for \u03b17 nAChR and cell viability was barely influenced by overexpression. While intracellular levels of \u03b17 nAChR were as anticipated, plasma membrane insertion was low. The glycosylation digest revealed no appreciable N-glycosylation product. This study demonstrates a stable and functional cell line expressing \u03b17 nAChR, whose protein expression, function, and viability are not affected by the absence of NACHO. The reduced plasma membrane insertion of \u03b17 nAChR, combined with incorrect matured N-glycosylation at the Golgi apparatus, suggests a loss of recognition signal for lectin sorting.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[44],"resource_type":[75],"application_areas":[],"therapeutic_areas":[55,143],"target_categories":[337],"target_families":[360],"targets":[189],"resource_keyword":[],"class_list":["post-14809","resource_library","type-resource_library","status-publish","hentry","products_tags-patchliner","resource_type-publication","therapeutic_areas-cns","therapeutic_areas-assay-development-and-validation","target_categories-ligand-gated-ion-channels","target_families-nicotinic-acetylcholine-receptors","targets-achr-7"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library\/14809","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=14809"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/products_tags?post=14809"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_type?post=14809"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/application_areas?post=14809"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/therapeutic_areas?post=14809"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_categories?post=14809"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_families?post=14809"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/targets?post=14809"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_keyword?post=14809"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}