Publications using NanoBiT

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  • Last Post 02 November 2017
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alandrem posted this 03 October 2016

Oh-hashi et al., 2016. SOD1 dimerization monitoring using a novel split NanoLuc, NanoBit. Cell Biochemistry and Function.

The authors use NanoBiT to study dimerization of WT and mutant SOD1 and also use NanoBiT to investigate the role of protein-protein interactions in the pathogenesis of amyotrophic lateral sclerosis (ALS).

Find the paper here:

http://onlinelibrary.wiley.com/doi/10.1002/cbf.3222/abstract

 

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alandrem posted this 31 October 2016

Cannaert et al, 2016. Detection and activity profiling of synthetic cannabinoids with a newly developed bio-assay. Analytical Chemistry.

The authors used NanoBiT to build B-arrestin recruitment assays for two G-protein coupled receptors (CB1 and CB2) in order to create live cell cannabinoid reporter assays. The assays were used to compare activity of synthetic cannabinoids and their metabolites and were also used to screen urine for CB receptor activity.

You can find the paper here. Also check out the lead authors discussion thread, "Our experience with NanoBiT".

http://pubs.acs.org/doi/abs/10.1021/acs.analchem.6b02600

A new publication by these authors that demonstrates a very nice application of this NanoBiT assay when further developed into stable cell lines: http://pubs.acs.org/doi/abs/10.1021/acs.analchem.7b02552

 

alandrem posted this 01 February 2017

Bodle et al., 2017. Development of a bimolecular luminescence complementation assay for RGS: G protein interactions in cells. Analytical Biochemistry: 522, 1-17.

This paper describes another application of NanoBiT to the study of GPCR signaling. The authors use NanoBiT to build a series of assays for the interaction of 15 different Regulator of G protein Signaling (RGS) proteins with different G protein subunits. The authors describe the process used to develop the assays, characterization of various RGS:G protein interactions, and also assess suitability for RGS screening assays.

You can find the paper here:

http://www.sciencedirect.com/science/article/pii/S0003269717300350

alandrem posted this 26 May 2017

Koushyar S. et al, 2017. The prohibitin-repressive interaction with E2F1 is rapidly inhibited by androgen signalling in prostate cancer cells. Oncogenesis (2017) 6, e333; doi:10.1038/oncsis.2017.32

The authors are working to better understand the development of androgen resistant prostate cancer, specifically the role of the androgen receptor corepressor, prohibitin (PHB), which is often downregulated by androgen treatment.  The authors identify PHB-mediated repression of genes essential for DNA replication and synthesis, (e.g. MCMs and TK1), via an E2F1 regulated pathway and further study the role of direct interaction between the PHB and E2F1 proteins using several methods: co-immunoprecipitation from prostate cancer nuclear extracts, Checkmate mammalian two-hybrid assay and a NanoBiT PPI assay that they developed. The authors performed the NanoBiT protein interaction assay (LgBiT-PHB, SmBiT-E2F1) in both COS-7 and LNCaP cells so that the interaction could be monitored in real-time in the presence of absence of androgen treatment and demonstrated low constitutive interaction which was inhibited by androgen treatment.

The article can be found here:  http://www.nature.com/oncsis/journal/v6/n5/full/oncsis201732a.html?WT.feed_name=subjects_cancer

 

 

alandrem posted this 08 June 2017

Nishiguchi, G. et al., 2017. Design and Discovery of N-(2-methyl-5'-morpholino-6'-((tetrahydro-2Hpyran-4-yl)oxy)-[3,3'-bipyridin]-5-yl)-3-(trifluoromethyl)benzamide (RAF709): A potent, selective and efficacious RAF inhibitor targeting RAS mutant cancers. J. Med. Chem., DOI: 10.1021/acs.jmedchem.6b01862

The major goal of this study was to identify selective RAF inhibitors that would suppress the RAF-MEK-ERKK pathway. A NanoBiT cRAF:bRAF interaction assay was used to study the potency of identified compounds to stabilize this dimer using dose-response assays in transfected HCT116 cells.

Here is the paper:

 http://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.6b01862

alandrem posted this 02 October 2017

Raj G. V. et al 2017. Estrogen receptor coregulator binding modulators (ERXs) effectively target estrogen receptor positive human breast cancers. eLife. 2017; 6: e26857.  http://dx.doi.org/10.7554/eLife.26857.001

The authors are studying synthetic peptides (peptidomimetics) and their ability to modulate estrogen receptor signaling. The authors developed NanoBiT PPI assays to study estrogen receptor and androgen receptor dimerization. Using these assays they were able to demonstrate compound ERX-11 blocks estrogen receptor signaling and can specifically disrupt ER dimerization, but does not affect AR dimerization. The authors also developed NanoBiT assay to study the interaction of ER with the co-regulator PELP1 and found this interaction could also be decreased with exposure to ERX-11.

GergoGogl posted this 30 October 2017

Hi,

I would like to advertise our article "Dynamic control of RSK complexes by phosphoswitch-based regulation", which was recently accepted at FEBS journal (doi=10.1111/febs.14311). We used Nanobit to detect the dissociation of ERK2 and RSK1 or MAGI1 and RSK1 in cells after EGF stimulation.

http://onlinelibrary.wiley.com/doi/10.1111/febs.14311/full

-We used structure based mutations as controls.
-As there was limited change in the signal we had to use corrections to eliminate the effect of substrate decay. We used 2 different normalization.

I think that it is a really nice example which shows the advantages of NanoBit compared to other cell based PPI assays.

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  • alandrem
alandrem posted this 01 November 2017

Thanks for sharing your recent publication! It is very interesting to see the rapid dynamics of this signaling pathway in your live cell model. We are actively working on some new substrates that provide a more stable signal. Although you were able to correct for the signal decay, these new substrates may be of interest to you. Let me know if you would like to evaluate in your model.

GergoGogl posted this 02 November 2017

Hi, Yes, it would be great to try a more stable substrate! We have a few other working NanoBiT constructs and sometimes (mostly in cases where the induced change of the signal is small) it is hard to correct to the spontaneous decay.

alandrem posted this 02 November 2017

Great. We are eager to get some feedback on the new substrates.  I will follow up with you to get your shipping information so that we can get some out to you.

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