Luminescence imaging has gained attention as a promising bio-imaging modality in situations where fluorescence imaging cannot be applied. However, wider application to multicolour and dynamic imaging is limited by the lack of bright luminescent proteins with emissions across the visible spectrum. Here we report five new spectral variants of the bright luminescent protein, enhanced Nano-lantern (eNL), made by concatenation of the brightest luciferase, NanoLuc, with various colour hues of fluorescent proteins. eNLs allow five-colour live-cell imaging, as well as detection of single protein complexes and even single molecules. We also develop an eNL-based Ca2+ indicator with a 500% signal change, which can image spontaneous Ca2+ dynamics in cardiomyocyte and neural cell models. These eNL probes facilitate not only multicolour imaging in living cells but also sensitive imaging of a wide repertoire of proteins, even at very low expression levels.
a b s t r a c tMore than 100 transthyretin (TTR) variants are associated with hereditary amyloidosis. Approaches for TTR amyloidosis that interfere with any step of the cascade of events leading to fibril formation have therapeutic potential. In this study we tested (À)-epigallocatechin-3-gallate (EGCG), the most abundant catechin of green tea, as an inhibitor of TTR amyloid formation. We demonstrate that EGCG binds to TTR ''in vitro" and ''ex vivo" and that EGCG inhibits TTR aggregation ''in vitro" and in a cell culture system. These findings together with the low toxicity of the compound raise the possibility of using EGCG in a therapeutic approach for familial amyloidotic polyneuropathy, the most frequent form of hereditary TTR amyloidosis.
Structured summary:MINT-7294529: TTR (uniprotkb:P02766) and TTR (uniprotkb:P02766) bind (MI:0407) by comigration in non-denaturing gel electrophoresis (MI:0404)
Microcalorimetric measurements have been made on the series of gemini surfactants [CMH2M+1(CH3)2N(CH2)SN(CH3)2CMH2M+1] Br2, designated CMCSCMBr2, where M and S indicate the numbers of carbons in the
side chains and spacer respectively, for M = 12 and S = 3, 4, 6, 8, 10, 12. For comparison, parallel
measurements have been made on the series of double-chain, singly charged surfactants dodecyldimethylalkylammonium bromides, designated as C12CNBr with N = 1, 2, 4, 6, 8, 10, 12, where N is the number of
carbons in the secondary alkyl chain. The choice of this second series was to compare each CMCSCMBr2 with
its nearest equivalent monomer C12CS/2Br. The values of the critical micelle concentrations (cmc) were found
to be in good agreement with other measurements, showing a maximum at S = 4−6 for the geminis and a
nonlinear variation with chain length for the C12CNBr series. For both series of surfactants, the enthalpies of
micellization, ΔH
mic, are all exothermic and show a marked minimum in magnitude at S = 4−6 or N = 4−6.
The variation of ΔH
mic and ΔS
mic across both series shows that the balance between enthalpic and entropic
contributions to the process of micellization changes substantially with S or N.
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