ArticleMolecular biotechnology2018
Application of NanoBiT for Monitoring Dimerization of the Null Hong Kong Variant of α-1-Antitrypsin, NHK, in Living Cells.
Article in Molecular biotechnology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 9 citations in OpenAlex.
- DNA Origami-Based Luminescent Biosensors Enabling Smartphone Detection of Nucleic Acid Sequences.ACS omega · 2025Article
- NanoBiT-based Analysis of Canine SOD1 Protein Dynamics: Understanding the Role of CCS and Ebselen Derivatives as Potential Therapeutics for Canine Degenerative Myelopathy.Cell biochemistry and biophysics · 2025Article
- Molecular characterization of the ER stress-inducible factor CRELD2.Cell biochemistry and biophysics · 2024Article
- Lumit: A Homogeneous Bioluminescent Immunoassay for Detecting Diverse Analytes and Intracellular Protein Targets.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Characterization of IRE1α in Neuro2a cells by pharmacological and CRISPR/Cas9 approaches.Molecular and cellular biochemistry · 2020Article
- A homogeneous bioluminescent immunoassay to probe cellular signaling pathway regulation.Communications biology · 2020Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
In this study, we investigated expression and dimerization of an ER-associated degradation (ERAD) substrate, a null Hong Kong variant of α-1-antitrypsin (NHK) using immunoblotting assay and a novel NanoLuc complementary reporter system called the NanoBiT (NB) assay. This NB-tagged NHK made it possible to monitor the intra- and extracellular status of NHK in living cells. The values for this NB assay fluctuated in response to distinct pharmacological stimuli and co-transfection of several ERAD-related factors. We then focused on mesencephalic astrocyte-derived neurotrophic factor (MANF), an unclarified ATF6/IRE1-downstream target, and established MANF-deficient Neuro2a (N2a) cells using CRISPR/Cas9 system. MANF-deficient N2a significantly elevated OS-9 protein after tunicamycin treatment; however, no specific differences in intra- and extracellular status of NHK protein were observed between wild-type and MANF-deficient cells. Taken together, intrinsic MANF in N2a cells is not strongly associated with the accumulation and clearance of unfolded proteins within the ER under current condition, but this novel NB assay is a useful approach for characterizing the protein status including ERAD substrates.
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