Chemical ubiquitination is an effective approach for accessing structurally defined, atypical ubiquitin (Ub) chains that are difficult to prepare by other techniques. Herein, we describe a strategy ...that uses a readily accessible premade isopeptide‐linked 76‐mer (isoUb), which has an N‐terminal Cys and a C‐terminal hydrazide, as the key building block to assemble atypical Ub chains in a modular fashion. This method avoids the use of auxiliary‐modified Lys and instead employs the canonical and therefore more robust Cys‐based native chemical ligation technique. The efficiency and capacity of this isoUb‐based strategy is exemplified by the cost‐effective synthesis of several linkage‐ and length‐defined atypical Ub chains, including K27‐linked tetra‐Ub and K11/K48‐branched tri‐, tetra‐, penta‐, and hexa‐Ubs.
All together now: Chemical protein synthesis enabled the generation of homogeneous atypical ubiquitin (Ub) chains that are difficult to prepare by other approaches. A ligation method mediated by an isopeptide‐linked Ub isomer was developed to assemble the Ub chains. In this approach, a premade isopeptide‐linked 76‐mer (isoUb) bearing an N‐terminal Cys and a C‐terminal hydrazide is the key building block.
Successive peptide ligation using a one‐pot method can improve the efficiency of protein chemical synthesis. Although one‐pot three‐segment ligation has enjoyed widespread application, a robust ...method for one‐pot four‐segment ligation had to date remained undeveloped. Herein we report a new one‐pot multisegment peptide ligation method that can be used to condense up to four segments with operational simplicity and high efficiency. Its practicality is demonstrated by the one‐pot four‐segment synthesis of a plant protein, crambin, and a human chemokine, hCCL21.
Pick up a peptide: Tfacm‐protected cysteine is readily activated by pH adjustment, enabling the development of a highly efficient one‐pot four‐segment ligation method. Two proteins, crambin and the chemokine hCCL21, are prepared using this rapid and high‐yielding synthetic route.
This paper examines the impact of a carbon trading pilot policy on the low-carbon international competitiveness of an industry to test whether creating a carbon market causes the Porter effect. Using ...a sample of 33 industries in 30 provinces in China from 2009 to 2016 with a difference-in-difference-in-difference model (DDD) and a series of robustness tests, we find evidence of a significant positive influence of a carbon trading pilot policy on the low-carbon international competitiveness of industries covered by the pilot programs. Research on its influencing mechanism reveals that a carbon trading pilot policy promotes the low-carbon international competitiveness of industries by driving low-carbon technological progress. Furthermore, a heterogeneity analysis of industry characteristics and carbon allowance allocation methods in different pilots indicates that the impact of a carbon trading pilot policy on industry low-carbon international competitiveness is reflected mainly in industries with low-carbon emissions, high state-owned capital, and high export intensity. Compared with the grandfather method and the historical intensity method, the low-carbon international competitiveness of an industry is significantly increased when the benchmarking method is used. The results of this paper offer important insights for improving the policy design of a nationwide carbon market, as well as a reference point for other countries and regions, especially developing countries, in establishing a carbon trading market.
China's pilot carbon trading policy is expected to be both efficient and flexible in reducing carbon emissions through incentivising low-carbon innovation. This paper analyses the effects of this ...pilot policy on low-carbon innovation using a difference-in-difference model, based on a sample of selected enterprises and carrying out a series of robustness tests to corroborate the results. The analysis shows that the pilot carbon trading policy is predicted to have a significantly positive effect on the low-carbon innovation of enterprises that fall under its scope, notably by alleviating obstacles to the financing of low-carbon innovation. Furthermore, a heterogeneity analysis of enterprises' characteristics and carbon allowance allocation methods in different pilots indicates that the effect of the pilot carbon trading policy on enterprises' low-carbon innovation will be reflected mainly in enterprises in China's eastern provinces, and in state-owned enterprises. Compared with the grandfathering and historical intensity allocation methods, the findings of this study suggest that the extent of low-carbon innovation is significantly greater when the benchmarking method is used. The results of this paper offer some key insights into improving the policy design of a nationwide carbon trading market in China, as well as a reference point for other countries and regions, especially developing countries, in establishing a carbon trading market.
Key policy insights
China's pilot carbon trading policy can promote low-carbon innovation.
Easing the financing constraints of enterprises can promote low-carbon innovation.
Compared with the grandfathering and historical intensity allocation methods, using the benchmarking method significantly improves the enterprises' low-carbon innovation.
Intratumoral heterogeneity is a hallmark of glioblastoma (GBM) tumors, thought to negatively influence therapeutic outcome. Previous studies showed that mesenchymal tumors have a worse outcome than ...the proneural subtype. Here we focus on STAT3 as its activation precedes the proneural-mesenchymal transition. We first establish a STAT3 gene signature that stratifies GBM patients into STAT3-high and -low cohorts. STAT3 inhibitor treatment selectively mitigates STAT3-high cell viability and tumorigenicity in orthotopic mouse xenograft models. We show the mechanism underlying resistance in STAT3-low cells by combining STAT3 signature analysis with kinome screen data on STAT3 inhibitor-treated cells. This allows us to draw connections between kinases affected by STAT3 inhibitors, their associated transcription factors and target genes. We demonstrate that dual inhibition of IGF-1R and STAT3 sensitizes STAT3-low cells and improves survival in mice. Our study underscores the importance of serially profiling tumors so as to accurately target individuals who may demonstrate molecular subtype switching.
As a flexible power grid load regulation facility, the orderly participation of electric vehicles (EVs) in power grid load regulation is conducive to the low-carbon transformation of energy system. ...To measure the co-benefit effect of orderly charging and discharging of EVs on carbon reduction, power grid security and economy, We build the calculation model of orderly charging and discharging of China’s EVs and compare the co-benefit effect of economic incentive orderly charging strategy, economic incentive + technology control orderly charging strategy under different penetration rates of EVs. Results show that in the Central China power grid, the peak load of can be reduced and valley load of can be improved by orderly charging and discharging with economic incentive + technology control strategy. With this method, 440 thousand EVs can reduce 31.2 thousand tons of coal consumption and 86.2 thousand tons of carbon emissions, save 4.29 billion yuan of project investment and increase 1.137 million KW grid margin. A 100% penetration rate of EVs can reduce 964.8 thousand tons of coal consumption and 2.25 million tons of carbon emissions, save 21.87 billion yuan of project investment, and increase grid margin by 5.79 million KW.
•Peak-valley price policy is not the most effective orderly charging strategy.•The co-benefit effect is the largest through economic incentive + technology control.•Control technology is more important than electricity price economic policy.
We report the chemical synthesis of the first photo-activatable protein antigen that can be used to study antigen-antibody interaction mediated responses in B cells. This strategy facilitated fine ...tuning of the caged protein antigen to optimize its bioactivity and photochemical properties. One optimal molecule, HEL-K
NPE, was totally inert to hen egg lysozyme (HEL)-specific B cells and could only restore its antigenicity upon photoactivation. Combined with real time live cell imaging, the utility of HEL-K
NPE was demonstrated as a proof of concept to quantify B cell synapse formation and calcium influx responses at the single cell level.
As an important mediator of information transfer between cells, exosomes play a unique role in regulating tumor growth, supporting vascular proliferation, tumor invasion, and metastasis. Exosomes are ...widely present in various body fluids, and therefore they can be used as a potential tool for non-invasive liquid biopsy. The present study reviews the role of exosomes in liquid biopsy, tumor microenvironment formation, and epithelial-mesenchymal transition in non-small cell lung cancer (NSCLC). By targeting epidermal growth factor receptor (EGFR) therapy as a first-line treatment for patients with NSCLC, this study also briefly describes the occurrence of EGRF+ exosomes and the role of exosomes and their contents in non-invasive detection and potential therapeutic targets in EGFR-mutated lung cancer.
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•A new one-pot ligation approach was developed for the synthesis of atypical Ub chains based on the Tfacm-isoUb building block.•The key intermidate Tfacm-isoUb building block can be ...readily prepared by Fmoc SPPS.•The practicality and efficiency of the new one-pot method is demonstrated by the successful preparation of K11-linked di-Ub.
Protein chemical synthesis can provide the homogeneous atypical ubiquitin chains that are usually difficult to obtain by other methods. Herein, we report a new one-pot ligation approach for the synthesis of atypical Ub chains based on the Tfacm-protected isoUb building block with operational simplicity and high efficiency. The key intermediate, the Tfacm-protected isoUb building block can be readily prepared by Fmoc SPPS. The practicality and efficiency of the new method is demonstrated by the successful preparation of K11-linked di-Ub.
The tomato leaf miner
(Meyrick) is one of the world's most destructive pests of tomato, and because of its severe economic impacts, as well as the development of pesticide resistance, the species has ...been intensively studied, especially in regard to the identification of targets for
control. However, functional genomic studies of
have been constrained by a lack of effective genetic tools. Therefore, the aim of the present study was to develop a CRISPR/Cas9 zygote microinjection protocol for generating heritable mutations in
, using the ommochrome synthesis gene
as an easily evaluated target gene. The injection of fertilised eggs with Cas9 protein and four sgRNAs, which targeted
exon 3, resulted in a mutagenesis rate of 31.9% for eggs reaching adulthood, and
mutagenesis resulted in either red or mosaic eye colour phenotypes. As such, this study is the first to report a complete and detailed CRISPR/Cas9 workflow for the efficient genome editing of the globally important invasive pest
. The application of this robust genome-editing tool to
will greatly facilitate the discovery of suitable RNAi control targets and the subsequent development of novel control strategies.
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