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Small ubiquitin-related modifier SUMO-3 is a member of a growing family of ubiquitin-like proteins (Ubls). So far, four isoforms of SUMO have been identified in humans. It is generally known that SUMO modification regulates protein localization and activity. Previous structure and function studies have been mainly focused on SUMO-1. The sequence of SUMO-3 is 46% identical with that of SUMO-1; nevertheless, functional heterogeneity has been found between the two homologues. Here we report the solution structure of SUMO-3 C47S (residues 14-92) featuring the beta-beta-alpha-beta-beta-alpha-beta ubiquitin fold. Structural comparison shows that SUMO-3 C47S resembles ubiquitin more than SUMO-1. On the helix-sheet interface, a strong hydrophobic interaction contributes to formation of the globular and compact fold. A Gly-Gly motif at the C-terminal tail, extending away from the core structure, is accessible to enzymes and substrates. In vivo, SUMO modification proceeds via a multistep pathway, and Ubc9 plays an indispensable role as the SUMO conjugating enzyme (E2) in this process. To develop a better understanding of SUMO-3 conjugation, the Ubc9 binding surface on SUMO-3 C47S has been detected by chemical shift perturbation using NMR spectroscopy. The binding site mainly resides on the hydrophilic side of the beta-sheet. Negatively charged and hydrophobic residues of this region are highly or moderately conserved among SUMO family members. Notably, the negatively charged surface of SUMO-3 C47S is highly complementary in its electrostatic potentials and hydrophobicity to the positively charged surface of Ubc9. This work indicates dissimilarities between SUMO-3 and SUMO-1 in tertiary structure and provides insight into the specific interactions of SUMO-3 with its modifying enzyme. 相似文献
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Construction of a recombinant BHV-1 expressing the VP1 gene of foot and mouth disease virus and its immunogenicity in a rabbit model 总被引:1,自引:0,他引:1
Xian-Gang Ren Fei Xue Yuan-Mao Zhu Guang-Zhi Tong Yan-Hui Wang Jun-Ke Feng Hong-Fei Shi Yu-Ran Gao 《Biotechnology letters》2009,31(8):1159-1165
Foot-and-mouth disease (FMD) and infectious bovine rhinotracheitis (IBR) are two important infectious diseases of cattle.
Using bovine herpesvirus type 1 (BHV-1) as a gene delivery vector for development of live-viral vaccines has gained widespread
interest. In this study, a recombinant BHV-1 was constructed by inserting the synthetic FMDV (O/China/99) VP1 gene in the
the gE locus of BHV-1 genome under the control of immediately early gene promoter of human cytomegalovirus (phIE CMV) and
bovine growth hormone polyadenylation (BGH polyA) signal. After homologous recombination and plaque purification, a recombinant
virus named BHV-1/gE−/VP1 was acquired and identified. The immunogenicity was confirmed in a rabbit model by virus neutralization test and enzyme-linked
immunosorbent assay (ELISA). The result indicated that the BHV-1/gE−/VP1 has the potential for being developed as a bivalent vaccine for FMD and IBR. 相似文献
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Ya-Ping Xue Wei Wang Ya-Jun Wang Zhi-Qiang Liu Yu-Guo Zheng Yin-Chu Shen 《Bioprocess and biosystems engineering》2012,35(9):1515-1522
To isolate enantioselective α-hydroxyacid dehydrogenases (α-HADHs), a high-throughput screening method was established. 2,4-Dinitrophenylhydrazine solution forms a red-brown complex with ketoacid produced during the α-HADH-mediated oxidation of α-hydroxyacid. The complex can be easily quantified by spectrophotometric measurement at 458?nm. The enantioselectivity of α-HADH in each strain can be measured with this colorimetric method using (R)- and (S)-α-hydroxyacid concurrently as substrates to evaluate the apparent enantioselectivity (E app). The E app closely matches the value of true enantioselectivity (E true) determined by HPLC analysis. With this method, a total of 34 stains harboring enantioselective α-HADHs were selected from 526 potential α-HADH-producing microorganisms. Pseudomonas aeruginosa displayed the highest (S)-enantioselective α-HADH activity. This strain appears promising for potential application in industry to produce (R)-α-hydroxyacids. The method described herein represents a useful tool for the high-throughput isolation of enantioselective α-HADHs. 相似文献
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Yuanyuan Yu Lei Hou Xu Shi Xiaoru Sun Xinhui Liu Yifan Yu Zhongshang Yuan Hongkai Li Fuzhong Xue 《PLoS genetics》2022,18(3)
Nonrandom selection in one-sample Mendelian Randomization (MR) results in biased estimates and inflated type I error rates only when the selection effects are sufficiently large. In two-sample MR, the different selection mechanisms in two samples may more seriously affect the causal effect estimation. Firstly, we propose sufficient conditions for causal effect invariance under different selection mechanisms using two-sample MR methods. In the simulation study, we consider 49 possible selection mechanisms in two-sample MR, which depend on genetic variants (G), exposures (X), outcomes (Y) and their combination. We further compare eight pleiotropy-robust methods under different selection mechanisms. Results of simulation reveal that nonrandom selection in sample II has a larger influence on biases and type I error rates than those in sample I. Furthermore, selections depending on X+Y, G+Y, or G+X+Y in sample II lead to larger biases than other selection mechanisms. Notably, when selection depends on Y, bias of causal estimation for non-zero causal effect is larger than that for null causal effect. Especially, the mode based estimate has the largest standard errors among the eight methods. In the absence of pleiotropy, selections depending on Y or G in sample II show nearly unbiased causal effect estimations when the casual effect is null. In the scenarios of balanced pleiotropy, all eight MR methods, especially MR-Egger, demonstrate large biases because the nonrandom selections result in the violation of the Instrument Strength Independent of Direct Effect (InSIDE) assumption. When directional pleiotropy exists, nonrandom selections have a severe impact on the eight MR methods. Application demonstrates that the nonrandom selection in sample II (coronary heart disease patients) can magnify the causal effect estimation of obesity on HbA1c levels. In conclusion, nonrandom selection in two-sample MR exacerbates the bias of causal effect estimation for pleiotropy-robust MR methods. 相似文献
40.
Effects of Melatonin on Growth,Physiology and Gene Expression in Rice Seedlings Under Cadmium Stress
Xiachen Lv Yunxia Fang Lantian Zhang Weiyi Zhang Ling Xu Jingjin Han Bailing Jin Xian Zhang Xiaoqin Zhang Dawei Xue 《Phyton》2019,88(2):91-100
Melatonin (MLT) is a hormonal substance found in many organisms and can improve plant stress resistance. In this study, the japonica rice variety Y32 and indica rice variety NJ6 were cultivated in hydroponics under different concentrations of CdCl2 at the two-leaf stage. The growth, physiological and biochemical responses of the seedlings and the expression of cadmium (Cd)-related genes under exogenous melatonin (MLT) treatment were assessed. The results indicated that Cd stress destroyed the dynamic balance between reactive oxygen species (ROS) production and removal, resulting in ROS accumulation, membrane lipid peroxidation, and impaired growth and development. Following the application of exogenous MLT to rice seedlings, increases in plant biomass including both underground and above-ground areas were observed. MLT also scavenged the inhibition of superoxide dismutase (SOD) and peroxidase (POD) in a concentration dependent manner in response to Cd stress. Catalase (CAT) activity and malondialdehyde (MDA) expression also decreased following MLT treatment. Amongst the six Cd-related genes assessed, five genes were down-regulated and one was up-regulated in response to MLT treatment. Taken together, these data demonstrate that MLT improves the resilience of rice seedlings at the biochemical, physiological, and molecular levels, and diminishes the damage caused by Cd stress. 相似文献