Cloning, expression, purification and crystallization of NHR3 domain from acute myelogenous leukemia-related protein AML1-ETO.

Abstract:

:The t(8;21) translocation is one of the most frequent chromosome abnormalities in acute myeloid leukemia. This translocation creates a fusion between the acute myelogenous leukemia 1 (AML1, a transcription factor) gene on chromosome 21 and the eight-twenty-one (ETO, a zinc finger nuclear protein) gene on chromosome 8, leading to the repression of certain AML1 target genes. We cloned NHR3 domain coding fragment into vector pGEX-6p-1 using PCR and obtained recombinant plasmid pGEX-6p-1-NHR3, which can be induced to stably overexpress fusion protein in E. coli. Through the purification on GST affinity chromatography column and PreScission protease cleavage, a large amount of NHR3 protein with high purity was obtained. In order to avoid possible interference of some strong negative charged molecules, NHR3 protein was further purified by Mono Q anion exchange chromatography. The NHR3 crystals were grown with hanging drop/vapor diffusion method and the first crystals appeared after four weeks at 18 degrees in 0.2 M Tris-sodium citrate dihydrate, 0.1 M sodium cacodylate, pH 6.5, and 30% iso-propanol (V/V). ESI mass spectrum showed that the molecular weight of this domain was in agreement with its primary structure sequence prediction, and circular dichroism spectral data (190-250 nm) showed that NHR3 had a well-defined secondary structure of 25.9% alpha-helix, 23.2% random coil and 50.9% turn, which was consistent with GOV4 software prediction.

authors

Yang HT,Wu DH,Xue XY,Liang WX,Miao XY,Pang H,Chen SJ

doi

10.1093/abbs/36.8.566

subject

Has Abstract

pub_date

2004-08-01 00:00:00

pages

566-70

issue

8

eissn

1672-9145

issn

1745-7270

journal_volume

36

pub_type

杂志文章
  • miR-4530 inhibits the malignant biological behaviors of human glioma cells by directly targeting RTEL1.

    abstract::Human glioma is the most common primary brain tumor and is associated with high morbidity and mortality. Aberrant expressions of microRNAs (miRNAs) are involved in glioma progression. In the present study, we aimed to elucidate the roles of miR-4530 in the pathogenesis of gliomas. miR-4530 expression was examined in h...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmaa126

    authors: Wang T,Zhang Y,Cui B,Wang M,Li Y,Gao K

    更新日期:2020-12-29 00:00:00

  • LT(K63/R72), a new mutant of Escherichia coli heat-labile enterotoxin, exhibits characteristics more similar to LT(K63) than LT(R72).

    abstract::LT(K63), a non-toxic mutant and LT(R72), a low toxic mutant of E. coli heat-labile enterotoxin are frequently used mucosal adjuvants. In many cases, the adjuvanticity of LT(K63) is lower than that of LT(R72), but LT(K63), which induces a mixed Th1/Th2 response, exhibits a higher level of protection than LT(R72) which ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:

    authors: Feng Q,Yang J,Luo P,Zhang WJ,Zou QM

    更新日期:2005-02-01 00:00:00

  • Identification of a G-quadruplex forming sequence in the promoter of UCP1.

    abstract::G-quadruplexes are higher-order nucleic acid structures formed in G-rich sequences in DNA or RNA. G-quadruplexes are distributed in many locations in the human genome, including promoter regions, and are viewed as promising therapeutic targets. Uncoupling protein-1 (UCP1) is a mitochondrial thermogenic gene critical f...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmy059

    authors: Zhao Y,Uhler JP

    更新日期:2018-07-01 00:00:00

  • Carbon monoxide augments electrical signaling in cultured neural networks of hippocampal neurons partly through activation of BKCa channels.

    abstract::Carbon monoxide (CO) is often viewed as a lethal gas in light of its capacity to prevent oxygen uptake in hemoglobin; however, it also functions to regulate a variety of proteins and physiological processes. Here we show that CO is an important chemical cue, to which neurons respond strongly, and this response is then...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmv017

    authors: Zhao H,Liu B,Zhang Q,Wu X,Yu Y,Jesse FF,Li L

    更新日期:2015-05-01 00:00:00

  • Effect of ozone produced from antibody-catalyzed water oxidation on pathogenesis of atherosclerosis.

    abstract::Recent studies have suggested that antibodies can catalyze the generation of unknown oxidants including hydrogen peroxide (H2O2) and ozone (O3) from singlet oxygen (1O2) and water. This study is aimed to detect the effect of antibody-catalyzed water oxidation on atherosclerosis. Our results showed that both H2O2 and O...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2006.00181.x

    authors: Peng KJ,Huang YS,An LN,Han XQ,Zhang JG,Wang QL,Sun J,Wang SR

    更新日期:2006-06-01 00:00:00

  • New methods for determining proton pumping ability and electron transfer activity of the cytochrome bc1 complex.

    abstract::It is important to determine the electron transfer activity and proton pumping ability of the cytochrome bc1 complex for better understanding its structure and function. In this study, several methods for determining the electron transfer and proton pumping of the bc1 complex, including the traditional and the new met...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmu126

    authors: Qu Y,Dong F

    更新日期:2015-02-01 00:00:00

  • Intein-mediated rapid purification of recombinant human pituitary adenylate cyclase activating polypeptide.

    abstract::In order to obtain the recombinant human PACAP efficiently by intein-mediated single column purification, a gene encoding human PACAP was synthesized and cloned into Escherichia coli expression vector pKYB. The recombinant vector pKY-PAC was transferred into E. coli ER2566 cells and the target protein was over-express...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/36.11.759

    authors: Yu RJ,Hong A,Dai Y,Gao Y

    更新日期:2004-11-01 00:00:00

  • Functional characterization of a special thermophilic multifunctional amylase OPMA-N and its N-terminal domain.

    abstract::A gene encoding a special thermophilic multifunctional amylase OPMA-N was cloned from Bacillus sp. ZW2531-1. OPMA-N has an additional 124-residue N-terminal domain compared with typical amylases and forms a relatively independent domain with a β-pleated sheet and random coil structure. Here we reported an unusual subs...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmr013

    authors: Li F,Zhu X,Li Y,Cao H,Zhang Y

    更新日期:2011-04-01 00:00:00

  • Argininosuccinate synthase 1 suppresses cancer cell invasion by inhibiting STAT3 pathway in hepatocellular carcinoma.

    abstract::Metastasis is the main reason for high recurrence and poor survival of hepatocellular carcinoma (HCC). The molecular mechanism underlying HCC metastasis remains unclear. In this study, we found that argininosuccinate synthase 1 (ASS1) expression was significantly decreased and down-regulation of ASS1 was closely corre...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmz005

    authors: Tao X,Zuo Q,Ruan H,Wang H,Jin H,Cheng Z,Lv Y,Qin W,Wang C

    更新日期:2019-03-01 00:00:00

  • Histone deacetylase SIRT6 inhibits glioma cell growth through down-regulating NOTCH3 expression.

    abstract::Gliomas are the most common brain tumors of the central nervous system. In this study, we investigated the molecular mechanisms and biological function of SIRT6 in human gliomas. The expression levels of SIRT6 in glioma tissues and cells were analyzed by qRT-PCR and western blot analysis. CCK8 and clonogenicity assays...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmy019

    authors: Chen X,Li D,Gao Y,Cao Y,Hao B

    更新日期:2018-04-01 00:00:00

  • Activation of liver X receptors promotes inflammatory cytokine mRNA degradation by upregulation of tristetraprolin.

    abstract::Liver X receptors (LXRs) have anti-inflammatory properties. Whether LXRs play a role in post-transcriptional control of inflammatory cytokine expression is not clear. Here, we firstly identified that the synthetic LXR agonist T0901317 promoted IL-1β, IL-6 and TNFα mRNA degradation. Moreover, T0901317 destabilized TNFα...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw136

    authors: Xiao J,Chen Q,Tang D,Ou W,Wang J,Mo Z,Tang C,Peng L,Wang D

    更新日期:2017-03-01 00:00:00

  • Cloning, sequence analysis and identification of a nonsense mutation-mediated mRNA decay of porcine GSTM2 gene.

    abstract::The glutathione S-transferase mu 2 gene (GSTM2) encodes a GST functioning in the elimination of electrophilic compounds and the regulation of cell growth. In this study, the sequence of porcine GSTM2 gene that contains the complete sequence encoding a protein of 218 amino acids was cloned. The deduced amino acid seque...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2007.00319.x

    authors: Huang J,Xiong Y,Deng C,Zuo B,Xu D,Lei M,Jiang S

    更新日期:2007-08-01 00:00:00

  • Correlation between circulating endothelial progenitor cells and serum carcinoembryonic antigen level in colorectal cancer.

    abstract::Circulating endothelial progenitor cells (cEPCs) play an important role in cancer development. Previous studies showed that serum carcinoembryonic antigen (CEA) levels and the number of circulating endothelial progenitor cells (cEPCs) in the peripheral blood are both involved in tumor neoangiogenesis, and can be used ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmx147

    authors: Li Y,Liu J,Zhao Z,Wen L,Li H,Ren J,Liu H

    更新日期:2018-03-01 00:00:00

  • Purification and partial characterization of beta-glucosidase from fresh leaves of tea plants (Camellia sinensis (L.) O. Kuntze).

    abstract::beta-Glucosidases are important in the formation of floral tea aroma and the development of resistance to pathogens and herbivores in tea plants. A novel beta-glucosidase was purified 117-fold to homogeneity, with a yield of 1.26%, from tea leaves by chilled acetone and ammonium sulfate precipitation, ion exchange chr...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2005.00053.x

    authors: Li YY,Jiang CJ,Wan XC,Zhang ZZ,Li DX

    更新日期:2005-06-01 00:00:00

  • Cloning and soluble expression of mature alpha-luffin from Luffa cylindrica and its antitumor activities in vitro.

    abstract::Luffin-a, a single-chain Type I ribosome-inactivating protein, which is known to be the most toxic of the luffin family and apparently possesses antitumor activity, was isolated from Luffa cylindrica seeds. In the present study, mature alpha-luffin was cloned from L. cylindrica and it was found that mature alpha-luffi...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmq056

    authors: Liu L,Wang R,He W,He F,Huang G

    更新日期:2010-08-01 00:00:00

  • Effects of raloxifene on caveolin-1 mRNA and protein expressions in vascular smooth muscle cells.

    abstract::Caveolin-1 is regulated by estrogen in vascular smooth muscle cells. Raloxifene, a selective estrogen receptor modulator that possibly has cardioprotective properties without an increased risk of cancer or other side effects of estrogen, may be used in women with risk of coronary artery disease. However, the relations...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2006.00222.x

    authors: Yang FL,He H,Liu XX,Tu B,Zeng XW,Su JX,Wang X,Hu Q

    更新日期:2006-11-01 00:00:00

  • Encapsulated paclitaxel nanoparticles exhibit enhanced anti-tumor efficacy in A549 non-small lung cancer cells.

    abstract::In the present study, paclitaxel (PTX) were encapsulated with polyethylene glycol (PEG)-polylactide (PLA)/D-α tocopheryl polyethylene glycol 1000 succinate (TPGS) (PEG-PLA/TPGS) and the enhanced anti-tumor activity of this PTX mixed micelles (PTX-MM) was evaluated in lung cancer cells. The PTX-MM prepared by a solvent...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmv110

    authors: Huang G,Zang B,Wang X,Liu G,Zhao J

    更新日期:2015-12-01 00:00:00

  • Construction of the seed-coat cDNA microarray and screening of differentially expressed genes in barley.

    abstract::Some barley mutants can synthesize neither anthocyanins nor proanthocyanidins in the seed coat, which is related to several genes in locus Ant13, but the exact model of action remains unknown. We used the cDNA microarray technology with barley transcription-deficient mutant (ant13-152) that does not synthesize proanth...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/36.10.695

    authors: Pang JS,He MY,Liu B

    更新日期:2004-10-01 00:00:00

  • Optimized adaptor polymerase chain reaction method for efficient genomic walking.

    abstract::Genomic walking is one of the most useful approaches in genome-related research. Three kinds of PCR-based methods are available for this purpose. However, none of them has been generally applied because they are either insensitive or inefficient. Here we present an efficient PCR protocol, an optimized adaptor PCR meth...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2006.00194.x

    authors: Xu P,Hu RY,Ding XY

    更新日期:2006-08-01 00:00:00

  • AMPKα1 regulates Idh2 transcription through H2B O-GlcNAcylation during brown adipogenesis.

    abstract::AMP-activated protein kinase (AMPK) is indispensable for the development and maintenance of brown adipose tissue (BAT), and its activity is inhibited due to obesity. The isocitrate dehydrogenase 2 (IDH2) is a mitochondrial enzyme responsible for the production of α-ketoglutarate, a key intermediate metabolite integrat...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmaa136

    authors: Cao Y,Liu X,Zhao J,Du M

    更新日期:2021-01-12 00:00:00

  • Biochemical characterization and ligand-binding properties of trehalose-6-phosphate phosphatase from Mycobacterium tuberculosis.

    abstract::Trehalose-6-phosphate phosphatase (TPP) is an essential enzyme for growth of mycobacteria, which has been identified to be a potential anti-tuberculosis drug target. However, the biochemical and ligand-binding properties and the 3D structure of TPP remain unclear so far. In the present study, we expressed the recombin...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmt084

    authors: Shi L,Zhang H,Qiu Y,Wang Q,Wu X,Wang H,Zhang X,Lin D

    更新日期:2013-10-01 00:00:00

  • Sequestration of glyceraldehyde-3-phosphate dehydrogenase to aggregates formed by mutant huntingtin.

    abstract::Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been reported to interact with proteins containing the polyglutamine (polyQ) domain. The present study was undertaken to evaluate the potential contributions of the polyQ and polyproline (polyP) domains to the co-localization of mutant huntingtin (htt) and GAPDH. Ov...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2007.00352.x

    authors: Wu J,Lin F,Qin Z

    更新日期:2007-11-01 00:00:00

  • Lysine methylation: beyond histones.

    abstract::Posttranslational modifications (PTMs) of histone proteins, such as acetylation, methylation, phosphorylation, and ubiquitylation, play essential roles in regulating chromatin dynamics. Combinations of different modifications on the histone proteins, termed 'histone code' in many cases, extend the information potentia...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1093/abbs/gmr100

    authors: Zhang X,Wen H,Shi X

    更新日期:2012-01-01 00:00:00

  • Endothelial cells modified by adenovirus vector containing nine copies hypoxia response elements and human vascular endothelial growth factor as the novel seed cells for bone tissue engineering.

    abstract::Vascularization is one of the hotspots during the development of new therapeutic strategies for bone tissue engineering, which can alleviate hypoxic circumstance and prevent transplant failure. Vascular endothelial growth factor (VEGF) gene transfection using recombinant adenovirus (Ad) vector can effectively promote ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmx101

    authors: Song X,Shi L,Chen L,Liu X,Qu X,Wang K,Wei F

    更新日期:2017-11-01 00:00:00

  • The role of P2X7 receptor in ATP-mediated human leukemia cell death: calcium influx-independent.

    abstract::Activation of the P2X7 receptor leads to a rapid, bidirectional flux of cations, causing broad range of biological responses including cytotoxicity. However, the mechanism of P2X7-mediated cytotoxicity remains largely unexplored. In our previous study, the lack of P2X7-mediated calcium response under normal conditions...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmp016

    authors: Zhang X,Meng L,He B,Chen J,Liu P,Zhao J,Zhang Y,Li M,An D

    更新日期:2009-05-01 00:00:00

  • Raman spectroscopic analysis of the effect of ultraviolet irradiation on calf thymus DNA.

    abstract::Raman spectroscopy was used for the first time to detect the effect of independent UVA (ultraviolet-A: 320-400 nm) and UVB (ultraviolet-B: 280-320 nm) irradiation on the calf thymus DNA in aqueous solution. After both UVA and UVB irradiation for 1 h or 3 h, the damage to the conformation of DNA was moderate, but the r...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/37.1.39

    authors: Tang YL,Guo ZY

    更新日期:2005-01-01 00:00:00

  • Caspase family proteases and apoptosis.

    abstract::Apoptosis, or programmed cell death, is an essential physiological process that plays a critical role in development and tissue homeostasis. The progress of apoptosis is regulated in an orderly way by a series of signal cascades under certain circumstances. The caspase-cascade system plays vital roles in the induction...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1111/j.1745-7270.2005.00108.x

    authors: Fan TJ,Han LH,Cong RS,Liang J

    更新日期:2005-11-01 00:00:00

  • Cloning and isolation of a Conus cysteine-rich protein homologous to Tex31 but without proteolytic activity.

    abstract::We cloned and isolated a cysteine-rich protein, designated Mr30, from Conus marmoreus. Mr30 belongs to the cysteine-rich secretory protein family that is highly homologous to Tex31 previously obtained from Conus textile and reported as a protease responsible for processing of pro-conotoxins. Mr30, purified by a proced...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2008.00385.x

    authors: Qian J,Guo ZY,Chi CW

    更新日期:2008-02-01 00:00:00

  • Wogonoside inhibits tumor growth and metastasis in endometrial cancer via ER stress-Hippo signaling axis.

    abstract::Wogonoside, a bioactive flavonoid component derived from Scutellaria baicalensis Georgi, has been reported to inhibit tumor growth in mice bearing various types of cancer cells such as breast cancer, lung cancer, and leukemia cells. However, whether wogonoside could inhibit tumor growth of endometrial cancer has not b...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmz109

    authors: Chen S,Wu Z,Ke Y,Shu P,Chen C,Lin R,Shi Q

    更新日期:2019-11-18 00:00:00

  • Production of the polyclonal anti-human metallothionein 2A antibody with recombinant protein technology.

    abstract::Metallothionein 2A (MT2A) is a small stress response protein that can be induced by exposure to toxic metals. It is highly expressed in breast cancer cells. In this study, the cDNA encoding the human MT2A protein was expressed as glutathione S-transferase (GST) fusion protein in Escherichia coli. Recombinant MT2A prot...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2006.00167.x

    authors: Marikar FM,Sun QM,Hua ZC

    更新日期:2006-05-01 00:00:00