Photo-caged 2-butene-1,4-dial as an efficient, target-specific photo-crosslinker for covalent trapping of DNA-binding proteins.

Covalent trapping of DNA-binding proteins via photo-crosslinking is an advantageous method for studying DNA-protein interactions. However, traditional photo-crosslinkers generate highly reactive intermediates that rapidly and non-selectively react with nearby functional groups, resulting in low target-capture yields and high non-target background capture. Herein, we report that photo-caged 2-butene-1,4-dial (PBDA) is an efficient photo-crosslinker for trapping DNA-binding proteins. Photo-irradiation (360 nm) of PBDA-modified DNA generates 2-butene-1,4-dial (BDA), a small, long-lived intermediate that reacts selectively with Lys residues of DNA-binding proteins, leading in minutes to stable DNA-protein crosslinks in up to 70% yield. In addition, BDA exhibits high specificity for target proteins, leading to low non-target background capture. The high photo-crosslinking yield and target specificity make PBDA a powerful tool for studying DNA-protein interactions.

Enzymolysis-based RNA pull-down identifies YTHDC2 as an inhibitor of antiviral innate response.

The innate immune response must be terminated in a timely manner at the late stage of infection to prevent unwanted inflammation. The role of m6A-modified RNAs and their binding partners in this process is not well known. Here, we develop an enzymolysis-based RNA pull-down (eRP) method that utilizes the immunoglobulin G-degrading enzyme of Streptococcus pyogenes (IdeS) to fish out m6A-modified RNA-associated proteins. We apply eRP to capture the methylated single-stranded RNA (ssRNA) probe-associated proteins and identify YT521-B homology domain-containing 2 (YTHDC2) as the m6A-modified interferon ß (IFN-ß) mRNA-binding protein. YTHDC2, induced in macrophages at the late stage of virus infection, recruits IFN-stimulated exonuclease ISG20 (IFN-stimulated exonuclease gene 20) to degrade IFN-ß mRNA, consequently inhibiting antiviral innate immune response. In vitro and in vivo deficiency of YTHDC2 increases IFN-ß production at the late stage of viral infection. Our findings establish an eRP method to effectively identify RNA-protein interactions and add mechanistic insight to the termination of innate response for maintaining homeostasis.

Identification of immune-activating metabolite for enhancing T cell therapy of cancer.

In this Research Highlight, we summarized the results from a study by Wu et al. published in Nature Cell Biology which uncovered that asparagine (Asn), a non-essential amino acid in mammalians, is able to enhance the TCR-mediated activation and efficacy of CD8+ T cells towards tumour through lymphocyte-specific protein tyrosine kinase (LCK) signalling. This study provides insights into the physiological function of Asn in T cell activation and effector functions, showing the encouraging possibility of key metabolites for empowering cancer immunotherapy.

Cysteine-specific protein multi-functionalization and disulfide bridging using 3-bromo-5-methylene pyrrolones.

Many reagents have been developed for cysteine-specific protein modification. However, few of them allow for multi-functionalization of a single Cys residue and disulfide bridging bioconjugation. Herein, we report 3-bromo-5-methylene pyrrolones (3Br-5MPs) as a simple, robust, and versatile class of reagents for cysteine-specific protein modification. These compounds can be facilely synthesized via a one-pot mild reaction and they show comparable tagging efficiency but higher cysteine specificity than the maleimide counterparts. The addition of cysteine to 3Br-5MPs generates conjugates that are amenable to secondary addition by another thiol or cysteine, making 3Br-5MPs valuable for multi-functionalization of a single cysteine and disulfide bridging bioconjugation. The labeling reaction and subsequent treatments are mild enough to produce stable and active protein conjugates for biological applications.

A novel synthesis of oleophylic Fe2O3/polystyrene fibers by γ-Ray irradiation for the enhanced photocatalysis of 4-chlorophenol and 4-nitrophenol degradation.

Photocatalytic degradation by efficient and easy recyclable semiconductor-catalysts is an ideal way to solve the environmental problem. A series of Fe2O3/Polystyrene (Fe2O3/PS) composite fibers with hydrophobic property were obtained by the electrospinning and γ-Ray irradiation methods. The γ-Ray irradiation treatment not only formed steady micro-nano construction of nanoparticles and fiber, but also reduced the hydroxyl group on Fe2O3 surface. The high photocatalytic efficiencies of Fe2O3/PS fiber were discovered with the high content of 4-chlorophenol (4-CP) and 4-nitrophenol (4-NP), due to the synergetic effect of adsorption and degradation. The suitable adsorption capacity of Fe2O3/PS could promote the utilization of the generated hydroxyl radical(OH) to directly oxidize the adsorbed pollutant molecules. Additionally, the photocatalytic activities for 4-CP and 4-NP still reached 80% and 75% in the 6th cycling and the composite fiber exhibited the good recyclability, which has the application development prospect for wastewater treatment. The mechanism of 4-CP and 4-NP decomposition was verified. Hence, the gained results could provide some insights into phenol degradation over the multifunctional and efficient catalyst.

4'-C-Trifluoromethyl modified oligodeoxynucleotides: synthesis, biochemical studies, and cellular uptake properties.

Herein, we report the synthesis of 4'-C-trifluoromethyl (4'-CF3) thymidine (T4'-CF3) and its incorporation into oligodeoxynucleotides (ODNs) through solid-supported DNA synthesis. The 4'-CF3 modification leads to a marginal effect on the deoxyribose conformation and a local helical structure perturbation for ODN/RNA duplexes. This type of modification slightly decreases the thermal stability of ODN/RNA duplexes (-1 °C/modification) and leads to improved nuclease resistance. Like the well-known phosphorothioate (PS) modification, heavy 4'-CF3 modifications enable direct cellular uptake of the modified ODNs without any delivery reagents. This work highlights that 4'-CF3 modified ODNs are promising candidates for antisense-based therapeutics, which will, in turn, inspire us to develop more potent modifications for antisense ODNs and siRNAs.