Advancements in the Development of non-BET Bromodomain Chemical Probes

The bromodomain and extra terminal (BET) family of bromodomain-containing proteins (BCPs) have been the subject of extensive research over the past decade, resulting in a plethora of high-quality chemical probes for their tandem bromodomains. In turn, these chemical probes have helped reveal the profound biological role of the BET bromodomains and their role in disease, ultimately leading to a number of molecules in active clinical development. However, the BET subfamily represents just 8/61 of the known human bromodomains, and attention has now expanded to the biological role of the remaining 53 non-BET bromodomains. Rapid growth of this research area has been accompanied by a greater understanding of the requirements for an effective bromodomain chemical probe and has led to a number of new non-BET bromodomain chemical probes being developed. Advances since December 2015 are discussed, highlighting the strengths/caveats of each molecule, and the value they add toward validating the non-BET bromodomains as tractable therapeutic targets.     

Discovery of Affinity-Based Probes for Btk Occupancy Assays

Bruton's tyrosine kinase (Btk) is an attractive target for the treatment of a wide array of B-cell malignancies and autoimmune diseases. Small-molecule covalent irreversible Btk inhibitors targeting Cys481 have been developed for the treatment of such diseases. In clinical trials, probe molecules are required in occupancy studies to measure the level of engagement of the protein by these covalent irreversible inhibitors. The result of this pharmacodynamic (PD) activity provides guidance for appropriate dosage selection to optimize inhibition of the drug target and correlation of target inhibition with disease treatment efficacy. This information is crucial for successful evaluation of drug candidates in clinical trials. Based on the pyridine carboxamide scaffold of a novel solvent-accessible pocket (SAP) series of covalent irreversible Btk inhibitors, we successfully developed a potent and selective affinity-based biotinylated probe 12 (2-[(4-{4-[5-(1-{5-[(3aS,4S,6aR)-2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-3,6,9,12-tetraoxapentadecan-15-amido)pentanoyl]piperazine-1-carbonyl}phenyl)amino]-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide). Compound 12 has been used in Btk occupancy assays for preclinical studies to determine the therapeutic efficacy of Btk inhibition in two mouse lupus models driven by TLR7 activation and type I interferon.     

Multi-functional biochip for medical diagnostics and pathogen detection

We describe a multi-functional biochip (MFB), which uses two different types of bioreceptors, including nucleic acid and antibody probes, on a single platform. The multi-functional capability of the MFB device for biomedical diagnostics is illustrated by measurements of DNA probes specific to gene fragments of Bacillus anthracis and antibody probes targeted to Escherichia coli. Calibration curves for monitoring pathogenic species using antibody probes against E. coli and DNA probes for B. anthracis illustrate the capability of the device for medical diagnostics and for quantitative detection of pathogenic agents.     

一种微机控制的多测头凸轮轴测量系统

本文介绍一种由机械装置，高精度圆光栅编码器，光栅位移传感器，微型计算机，接口控制电路等组成的多测头凸轮轴测量系统，该系统安装调整方便，定位准确，测量精度高，操作简单，工作可靠，能快速实现对凸轮轴的自动测量。     

Chemical Labeling of Protein 4′-Phosphopantetheinylation

Nature uses a diverse array of protein post-translational modifications (PTMs) to regulate protein structure, activity, localization, and function. Among them, protein 4′-phosphopantetheinylation derived from coenzyme A (CoA) is an essential PTM for the biosynthesis of fatty acids, polyketides, and nonribosomal peptides in prokaryotes and eukaryotes. To explore its functions, various chemical probes mimicking the natural structure of 4′-phosphopantetheinylation have been developed. In this minireview, we summarize these chemical probes and describe their applications in direct and metabolic labeling of proteins in bacterial and mammalian cells.     

N3-Kethoxal-Based Bioorthogonal Intracellular RNA Labeling

There is growing interest in developing intracellular RNA tools. Herein, we describe a strategy for N₃-kethoxal (N₃K)-based bioorthogonal intracellular RNA functionalization. With N₃K labeling followed by an in vivo click reaction with DBCO derivatives, RNA can be modified with fluorescent or phenol groups. This strategy provides a new way of labeling RNA inside cells.     

Study of secondary relaxations of poly(ethylene terephthalate) by photoluminescence technique

The α and β relaxation processes in two types of poly(ethylene terephthalate) with different degrees of crystallinity were studied by means of three methods, differential scanning calorimetry, dynamic-mechanical analysis and fluorescence spectroscopy. Information provided is complementary in the mean that every method sense phenomena that may occur at different times and length scales. Several probes, Coumarin 152 (C152), Coumarin 153 (C153), Coumarin 337 (C337) and 4′-dimethylamino-4-nitrostilbene (DMANS), were adsorbed in polymer films, and their fluorescence analysed over the temperature range from −150 to 150 °C. In general, a decrease in fluorescence intensity of probes as temperature increase was observed. This behaviour has been explained as a consequence of the enhancement of the free volume fraction that favoured the radiationless process of the lowest excited singlet state. Plots of fluorescence intensity versus temperature showed changes around the secondary relaxation temperatures. Therefore, good correlations between fluorescence and dynamic mechanical and calorimetric analysis were established. The obtained results indicated that the fluorescence from the probes incorporated to the material was dependent on the crystallinity of polymer. It would indicate that the fluorescence emission from those probes can be used to analyse annealing processes in semicrystalline polymers.     

Lanthanide-binding tags as versatile protein coexpression probes

Comprehensive proteomic analyses require new methodologies to accelerate the correlation of gene sequence with protein function. Key tools for such efforts include biophysical probes that integrate into the covalent architecture of proteins. Lanthanide-binding tags (LBTs) are expressible, multitasking fusion partners that are optimized to bind lanthanide ions and have several desirable attributes, which include long-lived luminescence, excellent X-ray scattering power for phase determination, and magnetic properties to facilitate NMR spectroscopic structure elucidation. Herein, we present peptide sequences with a 40-fold higher affinity for Tb(3+) ions and significantly brighter luminescence intensity compared with existing peptides. Incorporation of an LBT onto ubiquitin as a prototype fusion protein allows the use of powerful protein-visualization techniques, which include rapid luminescence detection of LBT-tagged proteins in SDS-PAGE gels, as well as determination of protein concentrations in complex mixtures. The LBT strategy is a new alternative for expressing fluorescent fusion proteins by routine molecular biological techniques.     

半导体纳米晶体制备及应用进展

综述了量子点的制备及应用进展