Reports : Aurora-A kinase-inactive mutants disrupt the interaction with Ajuba and cause defects in mitotic spindle formation and G2/M phase arrest in HeLa cells

( Mei Rong Bai ),( Jun Ni ),( Su Qin Shen ),( Qiang Huang ),( Jia Xue Wu ),( Yi Chen Le ),( Long Yu ) 생화학분자생물학회(구 한국생화학분자생물학회) 2014 BMB Reports Vol.47 No.11

Aurora-A is a centrosome-localized serine/threonine kinase that is overexpressed in multiple human cancers. We previously reported an intramolecular inhibitory regulation of Aurora-A between its N-terminal regulatory domain (Nt, amino acids [aa] 1-128) and the C-terminal catalytic domain (Cd, aa 129-403). Here, we demonstrate that although both Aurora-A mutants (AurA-K250G and AurA-D294G/Y295G) lacked interactions between the Nt and Cd, they also failed to interact with Ajuba, an essential activator of Aurora-A, leading to loss of kinase activity. Additionally, overexpression of either of the mutants resulted in centrosome amplification and mitotic spindle formation defects. Both mutants were also able to cause G2/M arrest and apoptosis. These results indicate that both K250 and D294/Y295 are critical for direct interaction between Aurora-A and Ajuba and the function of the Aurora-A complex in cell cycle progression. [BMB Reports 2014; 47(11): 631-636]

Enzyme-Catalyzed Henry Reaction in Choline Chloride-Based Deep Eutectic Solvents

( Xue Mei Tian ),( Suo Qin Zhang ),( Liang Yu Zheng ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.1

The enzyme-catalyzed Henry reaction was realized using deep eutectic solvents (DESs) as a reaction medium. The lipase from Aspergillus niger (lipase AS) showed excellent catalytic activity toward the substrates aromatic aldehydes and nitromethane in choline chloride:glycerol at a molar ratio of 1:2. Addition of 30 vol% water to DES further improved the lipase activity and inhibited DES-catalyzed transformation. A final yield of 92.2% for the lipase AS-catalyzed Henry reaction was achieved under optimized reaction conditions in only 4 h. In addition, the lipase AS activity was improved by approximately 3-fold in a DES.water mixture compared with that in pure water, which produced a final yield of only 33.4%. Structural studies with fluorescence spectroscopy showed that the established strong hydrogen bonds between DES and water may be the main driving force that affects the spatial conformation of the enzyme, leading to a change in lipase activity. The methodology was also extended to the aza-Henry reaction, which easily occurred in contrast to that in pure water. The enantioselectivity of both Henry and aza-Henry reactions was not found. However, the results are still remarkable, as we report the first use of DES as a reaction medium in a lipase-catalyzed Henry reaction.

Flavones from the Bark of Lindera caudata and Their Anti-Tobacco Mosaic Virus Activity

Yu-Chun Yang,Ying Qin,Xian-Xue Wu,Cong-Fang Xia,Yan-Lin Meng,Bin Zhou,Yan-Qing Ye,Xue-Mei Gao,Yin-Ke Li,Qiu-Fen Hu 대한화학회 2015 Bulletin of the Korean Chemical Society Vol.36 No.4

Two new flavones, 5-hydroxy-8-hydroxymethyl-7,4′-dimethoxy-flavone (1) and 6-hydroxy-8-hydroxymethyl-7,4′-dimethoxy-flavone (2), together with six known flavones (3–8), were isolated from the bark of Lindera caudata. The structures of 1–8 were elucidated by spectroscopic methods including extensive 1D and 2D NMR techniques. Compounds 1–8 were evaluated for their anti-tobacco mosaic virus (anti-TMV) activity. The results showed that Compounds 1 and 2 showed high anti-TMV activity with inhibition rates of 31.2 and 28.8%, respectively. These values are close to those of positive control.

Two new ursane-type triterpenoid saponins from Elsholtzia bodinieri

Jin-Dong Zhong,Xue-Wei Zhao,Xuan-Qin Chen,Hong-Mei Li,Chin-Ho Chen,Xue-Shan Xia,Rong-Tao Li 대한약학회 2016 Archives of Pharmacal Research Vol.39 No.6

Two new ursane-type triterpenoid saponins,bodiniosides M (1) and N (2), along with three knownsaponins, oblonganosides I (3), pseudobuxussaponin B (4)and bodinioside A (5), were isolated from the aerial partsof Elsholtzia bodinieri. The structures of compounds 1 and2 were characterized by spectroscopic data as well as acidhydrolysis and GC analysis as 3-O-b-D-xylopyranosyl-19ahydroxy-23-acetoxy-urs-12(13)-en-28-oic acid 28-O-a-Lrhamnopyranosyl-(1-2)-b-D-glucopyranoside and 3-O-b-Dglucopyranosyl-2a,19a-dihydroxy-urs-12(13)-en-28,20blactone. Compounds 1 and 5 exhibited potent anti-HCVactivities in vitro with a selective index of 6.53 and 4.41,respectively.

SPON2 Promotes M1-like Macrophage Recruitment and Inhibits Hepatocellular Carcinoma Metastasis by Distinct Integrin–Rho GTPase–Hippo Pathways

Zhang, Yan-Li,Li, Qing,Yang, Xiao-Mei,Fang, Fang,Li, Jun,Wang, Ya-Hui,Yang, Qin,Zhu, Lei,Nie, Hui-Zhen,Zhang, Xue-Li,Feng, Ming-Xuan,Jiang, Shu-Heng,Tian, Guang-Ang,Hu, Li-Peng,Lee, Ho-Young,Lee, Su-J American Association for Cancer Research 2018 Cancer research Vol.78 No.9

<P>Matricellular protein SPON2 acts as an HCC suppressor and utilizes distinct signaling events to perform dual functions in HCC microenvironment.</P><P>Tumor-associated macrophages (TAM) represent key regulators of the complex interplay between cancer and the immune microenvironment. Matricellular protein SPON2 is essential for recruiting lymphocytes and initiating immune responses. Recent studies have shown that SPON2 has complicated roles in cell migration and tumor progression. Here we report that, in the tumor microenvironment of hepatocellular carcinoma (HCC), SPON2 not only promotes infiltration of M1-like macrophages but also inhibits tumor metastasis. SPON2-α4β1 integrin signaling activated RhoA and Rac1, increased F-actin reorganization, and promoted M1-like macrophage recruitment. F-Actin accumulation also activated the Hippo pathway by suppressing LATS1 phosphorylation, promoting YAP nuclear translocation, and initiating downstream gene expression. However, SPON2-α5β1 integrin signaling inactivated RhoA and prevented F-actin assembly, thereby inhibiting HCC cell migration; the Hippo pathway was not noticeably involved in SPON2-mediated HCC cell migration. In HCC patients, SPON2 levels correlated positively with prognosis. Overall, our findings provide evidence that SPON2 is a critical factor in mediating the immune response against tumor cell growth and migration in HCC.</P><P><B>Significance:</B> Matricellular protein SPON2 acts as an HCC suppressor and utilizes distinct signaling events to perform dual functions in HCC microenvironment.</P><P><B>Graphical Abstract:</B> http://cancerres.aacrjournals.org/content/canres/78/9/2305/F1.large.jpg. <I>Cancer Res; 78(9); 2305–17. ©2018 AACR</I>.</P><P><B>Graphical Abstract</B></P><P> [Figure]</P>

Genome-wide analysis of codon usage bias patterns in an enterotoxigenic Escherichia coli F18 strain

Ri Wei Xia,Wen Bin Bao,Xue Mei Yin,Wei Yun Qin,Guo Qiang Zhu,Sheng Long Wu 한국유전학회 2017 Genes & Genomics Vol.39 No.11

Enterogenic Escherichia coli (ETEC) F18 strains are the main pathogenic bacteria causing severe diarrhea in humans and domestic animals. However, the information about synonymous codon usage pattern of ETEC F18 genome remains unclear. We conducted a genome-wide analysis of synonymous codon usage patterns in the ETEC F18 strain SRA: SAMN02471895. After filtering of the complete genome sequence, 4327 coding sequences were analyzed using multivariate statistical methods to calculate synonymous codon usage patterns and to evaluate the influence of various factors in shaping the codon usage. The mean GC content was 51.38%, with a slight preference for G/C-ending codons. Twenty-two codons were determined as ‘‘optimal codons”. ENC plots showed some of the genes were on or close to the expected curve, while only points with low-ENC values were below the curve. PR2 analysis showed that GC and AT were not used proportionally, suggesting major roles for mutational pressure and natural selection in shaping usage. Neutrality plots showed a significant correlation between GC12 and GC3, suggesting that mutational pressure is responsible for nucleotide composition in shaping the strength of codon usage. Translational selection was the main factor shaping the codon usage pattern of ETEC F18 genome, while other factors such as protein length, GRAVY and ARO values also influenced codon usage to some extent. We analyzed the codon usage pattern systematically and identified the factors shaping codon usage bias in the ETEC F18 genome. Such information further elucidates the mechanisms of synonymous codon usage bias and provides the basis of molecular genetic engineering and evolutionary studies.

ATAD2 is Highly Expressed in Ovarian Carcinomas and Indicates Poor Prognosis

Wan, Wei-Na,Zhang, Yi-Xia,Wang, Xue-Mei,Liu, Yan-Jun,Zhang, Yu-Qin,Que, Yan-Hong,Zhao, Wen-Jing Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

The purpose of this study was to explore the expression of ATAD2 in ovarian tumor tissue as well as its relationship with degree of malignancy. Tumor tissue from 110 cases of ovarian cancer was collected in accordance with the Declaration of Helsinki for evaluation of ATAD2 expression iimmunohistochemistry, quantitative PCR (qPCR) and Western blotting. The correlation between the ATAD2 expression and and the prognosis of ovarian cancer was evaluated by Cox regression model. In addition, HO-8910 and OVCAR-3 cells were transfected with two siRNAs targeting ATAD2. Cell viability was evaluated with MTT assay, and cell migration by transwell migration assay. ATAD2 was shown to be highly expressed in 65.5% (72/110) of ovarian cancer cases, both at transcriptional and protein levels. Moreover, highly expression was positively correlated with degree of malignancy. Knock-down of ATAD2 in HO-8910 and OVCAR-3 cells was found to reduce cell migration. In addition, follow-up visits of the patients demonstrated that the 5-year survival rate was lower in patients with high expression of ATAD2. Our study suggested that ovarian tumor tissue may have highly expressed ATAD2, which is associated with tumor stage, omentum-metastasis, ascites and CA-125. Increased ATAD2 may play important roles in tumor proliferation and migration. ATAD2 could serve in particular as a prognostic marker and a therapeutic target for ovarian cancer.

Species Classification and Quality Assessment of Chaihu (Radix Bupleuri) Based on High-Performance Liquid Chromatographic Fingerprint and Combined Chemometrics Methods

Xiao-Jie Li,Jie Hu,Zhen-Yu Li,Xue-Mei Qin,Li-Zeng Zhang,Xiao-Qing Guo 대한약학회 2011 Archives of Pharmacal Research Vol.34 No.6

A high-performance liquid chromatographic (HPLC) method was established to analyze 36 Chaihu (Radix Bupleuri) samples collected from three species (Bupleurum chinense DC., B. scorzonerifolium Willd. and B. smithii Wolff.). Addition of trifluoroacetic acid into the mobile phase resulted in fingerprint chromatograms with stable baselines. There were thirty-two characteristic peaks in the standard fingerprint of B. chinense DC. Different recognition pattern methods, including similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA) and partial least squares-discrimination analysis (PLS-DA) were utilized to analyze the 36 samples based on the contents of chemical constituents. Consistent results from SA, HCA and PCA analysis illustrated the rationalisation for why B. smithii Wolff. was not quoted in the Chinese Pharmacopoeia and classified samples were in agreement with their species. PLS-DA loading plots showed the chemical markers which had the most influences on the separation among different species. However, SA, HCA and PCA could not differentiate between wild and cultivated B. chinense DC. as well as between samples from different provinces. HPLC fingerprint in combination with chemometric techniques provided a very flexible and reliable method for homogeneity evaluation and quality assessment of traditional Chinese medicine.

Covalent Immobilization of Penicillin G Acylase onto Fe₃O₄@Chitosan Magnetic Nanoparticles

( Xiao Min Ling ),( Xiang Yu Wang ),( Ping Ma ),( Yi Yang ),( Jie Mei Qin ),( Xue Jun Zhang ),( Ye Wang Zhang ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.5

Penicillin G acylase (PGA) was immobilized on magnetic Fe₃O₄@chitosan nanoparticles through the Schiff base reaction. The immobilization conditions were optimized as follows: enzyme/support 8.8 mg/g, pH 6.0, time 40 min, and temperature 25℃. Under these conditions, a high immobilization efficiency of 75% and a protein loading of 6.2 mg/g-support were obtained. Broader working pH and higher thermostability were achieved by the immobilization. In addition, the immobilized PGA retained 75% initial activity after ten cycles. Kinetic parameters Vmax and Km of the free and immobilized PGAs were determined as 0.113 mmol/min/mg-protein and 0.059 mmol/min/mg-protein, and 0.68 mM and 1.19 mM, respectively. Synthesis of amoxicillin with the immobilized PGA was carried out in 40% ethylene glycol at 25℃ and a conversion of 72% was obtained. These results showed that the immobilization of PGA onto magnetic chitosan nanoparticles is an efficient and simple way for preparation of stable PGA.

Butyrolactones Derivatives from the Fermentation Products of an Endophytic Fungus Aspergillus versicolor

Ye, Yan-Qing,Xia, Cong-Fang,Yang, Juan-Xia,Yang, Yu-Chun,Qin, Ying,Gao, Xue-Mei,Du, Gang,Li, Xue-Mei,Hu, Qiu-Fen Korean Chemical Society 2014 Bulletin of the Korean Chemical Society Vol.35 No.10

Two new butyrolactones, asperphenol A (1) and B (2), together with four known butyrolactones (3-6) were isolated from the fermentation products of an endophytic fungus Aspergillus versicolor. Their structures were elucidated by spectroscopic methods including extensive 1D- and 2D-NMR techniques. Compounds 1-6 were also tested for their anti-tobacco mosaic virus (anti-TMV) activities. The results showed that compound 2 exhibited high anti-TMV activity with inhibition rate of 46.7%. The other compounds also exhibited potential anti-TMV activities with inhibition rates in the range of 21.8-28.4%.