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115(25), E5776-E5785. ABC transporter content diversity in Streptococcus pneumoniae impacts competence regulation and bacteriocin production. 2018

Wang, C.Y., Patel, N., Wholey, W.Y., and Dawid, S.

Notes: S. pneumoniae uses the ABC transporters ComAB and BlpAB in natural competence and bacterial predation through bacteriocin secretion. NanoLuc and RFluc constructs with upstream comAB and blpAB promoters were used to look at transcriptional activation. Also, bacteriocin secretion by ComAB and BlpAB was determined using the HiBiT system. BlpI was tagged with HiBiT and luciferase activity was monitored in both wild-type and mutant transporter backgrounds. Both ComAB and BlpAB were able to respond to and secrete BlpI. (5137)

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Virus Res. 243, 69–74. Development of a rapid and quantitative method for the analysis of viral entry and release using the NanoLuc® luciferase complementation assay. 2017

Sasaki, M., Anindita, P.D., Phongphaew, W., Carr, M., Kobayashi, S., Orba, Y. and Sawa, H.

Notes: The authors developed quantitative methods for the detection of cellular entry and release of subviral and flavivirus-like particles (SVPs, VLPs) by tagging the particles with HiBiT. The HiBiT tag was used for quantitation of the particles, and cellular entry was studied using a LgBiT-expressing stable Vero cell line. (4930)

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Biochim. Biophys. Acta 1864, 2322–9. Regions of MRAP2 required for the inhibition of orexin and prokineticin receptor signaling. 2017

Rouault, A.A.J., Lee, A.A. and Sebag, J.A.

Notes: Authors demonstrate use of HiBiT extracellular system to study the role of MRAP2 in GPCR trafficking and surface expression. (4929)

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Biochem. Biophys. Rep. 12, 40–5. Application of a novel HiBiT peptide tag for monitoring ATF4 protein expression in Neuro2a cells. 2017

Oh-Hashi, K., Furuta, E., Fujimura, K. and Hirata, Y.

Notes: The authors used CRISPR/Cas9 editing to tag endogenous ATF4 with HiBiT and then studied changes in abundance following various cell treatments. (4928)

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ACS Chemical Biology Epub before print. CRISPR-Mediated Tagging of Endogenous Proteins with a Luminescent Peptide 2017

Schwinn, M.K., Machleidt, T., Zimmerman, K., Eggers, C.T., Dixon, A.S., Hurst, R., Hall, M.P., Encell, L.P., Binkowski, B.F. and Wood, K.V.

Notes: Using CRISPR/Cas9 gene editing, HiBiT was tagged to the C terminus of HIF1α and several of its downstream transcriptional target proteins. The Nano-Glo® Lytic Detection System was used to quantify HiBiT-tagged endogenous protein levels, and the Nano-Glo® Live Cell Assay System was used for real-time detection in live cells. Bioluminescence was detected using the GloMax® Discover System. This study demonstrated the ability to efficiently tag endogenous proteins with HiBiT, allowing fast and sensitive quantification of the response dynamics in their regulated expression and covalent modifications. (4869)

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Biochim. Biophys. Acta Epub before print. Regions of MRAP2 required for the inhibition of orexin and prokineticin receptor signaling 2017

Rouault, A.A.J., Lee, A.A. and Sebag, J.A.

Notes: The Nano-Glo® HiBiT Extracellular Detection System was used to detect surface density of receptor proteins. (4880)

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Sci. Rep. 6, 29130. Determination of GLUT1 oligomerization parameters using bioluminescent Förster resonance energy transfer. 2016

Looyenga, B., VanOpstall, C., Lee. Z., Bell, J., Lodge, E., Wrobel, K., Arnoys, E. and Louters, L.

Notes: Oligomerization of the glucose transporter (GLUT1) within this plasma membrane was assessed using the NanoBRET™ system. When expressed at high levels, GLUT1 has been shown to form tetrameric complexes with higher transport efficiency. Theoretical NanoBRET™ efficiency was determined for a variety of fluorescent protein acceptors and experiments were performed with mCherry as the NanoBRET™ acceptor. GLUT1 was shown to form a range of higher order complexes in live cells. Flow cytometry and immunoblotting were used in parallel to estimate GLUT1 density in cells. (5054)

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