In eukaryotic cells, the shortening and removal of the poly(A) tail of cytoplasmic mRNA by deadenylase enzymes is a critical step in post-transcriptional gene regulation. The ribonuclease activity of deadenylase enzymes is attributed to either a DEDD (Asp-Glu-Asp-Asp) or an endonuclease–exonuclease–phosphatase domain. Both domains require the presence of two Mg2+ ions in the active site. To facilitate the biochemical analysis of deadenylase enzymes, we have developed a fluorescence-based deadenylase assay. The assay is based on end-point measurement, suitable for quantitative analysis and can be adapted for 96- and 384-well microplate formats. We demonstrate the utility of the assay by screening a chemical compound library, resulting in the identification of non-nucleoside inhibitors of the Caf1/CNOT7 enzyme, a catalytic subunit of the Ccr4–Not deadenylase complex. These compounds may be useful tools for the biochemical analysis of the Caf1/CNOT7 deadenylase subunit of the Ccr4–Not complex and indicate the feasibility of developing selective inhibitors of deadenylase enzymes using the fluorescence-based assay.
Polymorphisms
in the region of the calmodulin-dependent kinase
isoform D (CaMK1D) gene are associated with increased incidence of
diabetes, with the most common polymorphism resulting in increased
recognition by transcription factors and increased protein expression.
While reducing CaMK1D expression has a potentially beneficial effect
on glucose processing in human hepatocytes, there are no known selective
inhibitors of CaMK1 kinases that can be used to validate or translate
these findings. Here we describe the development of a series of potent,
selective, and drug-like CaMK1 inhibitors that are able to provide
significant free target cover in mouse models and are therefore useful
as
in vivo
tool compounds. Our results show that
a lead compound from this series improves insulin sensitivity and
glucose control in the diet-induced obesity mouse model after both
acute and chronic administration, providing the first
in vivo
validation of CaMK1D as a target for diabetes therapeutics.
Abstract. In a simple reaction between Silver nitrate and Thiourea under the presence of Chitosan, novel fork architectures of silver sulfide (Ag 2 S) were obtained through In-situ self-assembly of
This paper reports functionalized zinc sulphide (ZnS) semiconductor nanocrystals (quantum dots, approx., 2.5 nm) which are an important building block in self-assembled nanostructures. ZnS is functionalized by organic stabilizer Thio glycolic acid (TGA). The samples have been synthesized by colloidal technique at relatively low temperature (below 100• C) at an atmospheric pressure of 10 −3 torr. Manganese (Mn) doping ions have been incorporated (doped) in ZnS host lattice and observed its effect on growth morphology and optical properties of ZnS colloidal nanocrystals. By XRD, SEM, TEM, and PL, the obtained cubic phase nanosized TGA-capped ZnS materials were characterized. The morphology of ZnS obtained at different temperatures are analyzed by SEM. The crystallite size of the ZnS nanoparticles was estimated from the X-ray diffraction pattern by using Scherrer's formula (approximately 2.5 nm) which is confirmed by TEM. The estimated bandgap value of ZnS NC's by (αhύ) 2 versus hύ plot was 4.89 eV. Gaussian fitting curve in photoluminescence (PL) spectra indicated room temperature emission wavelength range from 300 to 500 nm in undoped and Mn-doped ZnS, with different emission peak intensities, and suggested the wide band emission colours in visible and near UV region which has wider applications in optical devices.
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