دورية أكاديمية
N-Glycosylation Deficiency in Transgene α7 nAChR and RIC3 Expressing CHO Cells Without NACHO.
العنوان: | N-Glycosylation Deficiency in Transgene α7 nAChR and RIC3 Expressing CHO Cells Without NACHO. |
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المؤلفون: | Brockmöller S; Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany. Sabrina.Brockmoeller@yahoo.de., Molitor LM; Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany., Seeger T; Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany., Worek F; Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany., Rothmiller S; Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany. |
المصدر: | The Journal of membrane biology [J Membr Biol] 2024 Aug; Vol. 257 (3-4), pp. 245-256. Date of Electronic Publication: 2024 Jul 05. |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Springer Country of Publication: United States NLM ID: 0211301 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-1424 (Electronic) Linking ISSN: 00222631 NLM ISO Abbreviation: J Membr Biol Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: New York, Springer. |
مواضيع طبية MeSH: | alpha7 Nicotinic Acetylcholine Receptor*/genetics , alpha7 Nicotinic Acetylcholine Receptor*/metabolism , Cricetulus*, Animals ; CHO Cells ; Glycosylation ; Humans ; Cricetinae ; Transgenes ; Molecular Chaperones/genetics ; Molecular Chaperones/metabolism ; Cell Survival/genetics ; Intracellular Signaling Peptides and Proteins |
مستخلص: | The human neuronal nicotinic acetylcholine receptor α7 (nAChR) is an important target implicated in diseases like Alzheimer's or Parkinson's, as well as a validated target for drug discovery. For α7 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 α7 nAChR. This study investigates the consequence of NACHO absence on α7 nAChR expression and function. Therefore, the sequences of human α7 nAChR and human RIC3 were transduced in Chinese hamster ovary (CHO) cells. Protein expression and function of α7 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 α7 nAChR and cell viability was barely influenced by overexpression. While intracellular levels of α7 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 α7 nAChR, whose protein expression, function, and viability are not affected by the absence of NACHO. The reduced plasma membrane insertion of α7 nAChR, combined with incorrect matured N-glycosylation at the Golgi apparatus, suggests a loss of recognition signal for lectin sorting. (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.) |
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فهرسة مساهمة: | Keywords: N-Glycosylation; NACHO; Nicotinic acetylcholine receptor; RIC3 |
المشرفين على المادة: | 0 (alpha7 Nicotinic Acetylcholine Receptor) 0 (RIC3 protein, human) 0 (Molecular Chaperones) 0 (Intracellular Signaling Peptides and Proteins) |
تواريخ الأحداث: | Date Created: 20240705 Date Completed: 20240730 Latest Revision: 20240730 |
رمز التحديث: | 20240730 |
DOI: | 10.1007/s00232-024-00317-0 |
PMID: | 38967800 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1432-1424 |
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DOI: | 10.1007/s00232-024-00317-0 |