Biophysical characterization and ligand-binding properties of the elongation factor Tu from Mycobacterium tuberculosis.

Abstract:

:Mycobacterium tuberculosis (Mtb) is the key devastating bacterial pathogen responsible for tuberculosis. Increasing emergence of multi-drug-resistant, extensively drug-resistant, and rifampicin/isoniazid-resistant strains of Mtb makes the discovery of validated drug targets an urgent priority. As a vital translational component of the protein biosynthesis system, elongation factor Tu (EF-Tu) is an important molecular switch responsible for selection and binding of the cognate aminoacyl-tRNA to the acceptor site on the ribosome. In addition, EF-Tu from Mtb (MtbEF-Tu) is involved in the initial step of trans-translation which is an effective system for rescuing the stalled ribosomes from non-stop translation complexes under stress conditions. Given its crucial role in protein biosynthesis, EF-Tu is identified as an excellent molecular target for drug design. Here, we reported the recombinant expression, purification, biophysical characterization, and structural modeling of the MtbEF-Tu protein. Our results demonstrated that prokaryotic expression plasmids of pET28a-MtbEF-Tu could be expressed efficiently in Escherichia coli. We successfully purified the 6× His-tagged proteins with a yield of 16.8 mg from 1 l of Luria Bertani medium. Dynamic light scattering experiments showed that MtbEF-Tu existed in a monomeric form, and circular dichroism experiments indicated that MtbEF-Tu was well structured. Moreover, isothermal titration calorimetry experiments displayed that the purified MtbEF-Tu protein possessed intermediate binding affinities for guanosine-5'-triphosphate (GTP) and GDP. The GTP/GDP-binding sites were predicted by flexible molecular docking approach which reveals that GTP/GDP binds to MtbEF-Tu mainly through hydrogen bonds. Our work lays the essential basis for further structural and functional studies of MtbEF-Tu as well as MtbEF-Tu-related novel drug developments.

authors

Yang J,Hong J,Luo L,Liu K,Meng C,Ji ZL,Lin D

doi

10.1093/abbs/gmy164

subject

Has Abstract

pub_date

2019-02-01 00:00:00

pages

139-149

issue

2

eissn

1672-9145

issn

1745-7270

pii

5274658

journal_volume

51

pub_type

杂志文章
  • miR-375 is highly expressed and possibly transactivated by achaete-scute complex homolog 1 in small-cell lung cancer cells.

    abstract::In this study, we identified five miRNAs highly expressed in the small-cell lung cancer (SCLC) cell line NCI-H209. Among them, the expression levels of miR-375 were dramatically elevated in all SCLC cell lines examined, coincident with the expression of the transcription factor achaete-scute complex homolog 1 (ASCL1)....

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmr110

    authors: Zhao H,Zhu L,Jin Y,Ji H,Yan X,Zhu X

    更新日期:2012-02-01 00:00:00

  • Snapshot of the interaction between HIV envelope glycoprotein 120 and protein disulfide isomerase.

    abstract::The human immunodeficiency virus-1 (HIV-1) envelope glycoprotein 120 (gp120) binds to cell surface receptors and mediates HIV entry. Previous studies suggest the cell surface protein disulfide isomerase (PDI) might interact with disulfide bond(s) of gp120 and thus facilitate HIV-1 entry. In the present study, a kineti...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmq024

    authors: Wang Z,Zhou Z,Guo ZY,Chi CW

    更新日期:2010-05-15 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

  • L-amino acid oxidase from Naja atra venom activates and binds to human platelets.

    abstract::An L-amino acid oxidase (LAAO), NA-LAAO, was purified from the venom of Naja atra. Its N-terminal sequence shows great similarity with LAAOs from other snake venoms. NA-LAAO dose-dependently induced aggregation of washed human platelets. However, it had no activity on platelets in platelet-rich plasma. A low concentra...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2008.00372.x

    authors: Li R,Zhu S,Wu J,Wang W,Lu Q,Clemetson KJ

    更新日期:2008-01-01 00:00:00

  • The structure-function relationship of MSI7, a matrix protein from pearl oyster Pinctada fucata.

    abstract::We previously identified a matrix protein, MSI7, from pearl oyster Pinctada fucata. According to the structural analysis, the DGD site in the N-terminal of MSI7 is crucial for its role in the shell formation. In this study, we expressed a series of recombinant MSI7 proteins, including the wild-type and several mutants...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmp086

    authors: Feng Q,Fang Z,Yan Z,Xing R,Xie L,Zhang R

    更新日期:2009-11-01 00:00:00

  • Neuroprotective effect of deferoxamine on N-methyl-d-aspartate-induced excitotoxicity in RGC-5 cells.

    abstract::Many N-methyl-d-aspartate (NMDA) receptor antagonists have been used to treat neurodegenerative diseases induced by glutamate excitotoxicity in clinics. However, the universality of the glutamic acid neurotransmitter system makes the glutamic acid receptor blockers inefficient and unsafe. Thus, regulating the downstre...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmx082

    authors: Tian Y,He Y,Song W,Zhang E,Xia X

    更新日期:2017-09-01 00:00:00

  • Kaempferol inhibits the growth and metastasis of cholangiocarcinoma in vitro and in vivo.

    abstract::Kaempferol is a flavonoid that has been reported to exhibit antitumor activity in various malignant tumors. However, the role of kaempferol on cholangiocarcinoma (CCA) is largely unknown. In this article, we found that kaempferol inhibited proliferation, reduced colony formation ability, and induced apoptosis in HCCC9...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmv133

    authors: Qin Y,Cui W,Yang X,Tong B

    更新日期:2016-03-01 00:00:00

  • Small hairpin RNA targeting at Bcl-2 increases cytarabine-induced apoptosis in Raji cells.

    abstract::Bcl-2, a prominent member of the Bcl-2 family proteins, is responsible for the dysregulation of apoptosis and resistance to chemotherapy. In this study, we investigated whether small hairpin RNA (shRNA) targeting at Bcl-2 mRNA could enhance cytarabine (Ara-C)-induced apoptosis in Raji cells. Bcl-2 shRNA was transfecte...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmn015

    authors: He D,Fang B

    更新日期:2009-02-01 00:00:00

  • Identification and functional characterization of a novel missense mutation in FRMD7 responsible for idiopathic congenital nystagmus.

    abstract::Idiopathic congenital nystagmus (ICN) is a genetically heterogeneous eye movement disorder which seriously reduces childhood visual acuity. X-linked inheritance is the most common pattern, and mutations in FERM domain-containing protein 7 (FRMD7) are the major cause. Here, we recruited a four-generation Chinese family...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmy161

    authors: Wang Z,Wang M,Wang C,Lu B

    更新日期:2019-02-01 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

  • Neuronal conditional knockout of NRSF decreases vulnerability to seizures induced by pentylenetetrazol in mice.

    abstract::Neuron restrictive silencer factor (NRSF), also known as repressor element-1 silencing transcription factor, has been reported to modulate neuronal excitability and acts as endogenous anticonvulsant in kainic acid-induced or kindling-evoked seizure activity. However, whether NRSF functions in pentylenetetrazol (PTZ)-i...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gms023

    authors: Liu M,Sheng Z,Cai L,Zhao K,Tian Y,Fei J

    更新日期:2012-06-01 00:00:00

  • Heat shock protein 90 mediates the apoptosis and autophage in nicotinic-mycoepoxydiene-treated HeLa cells.

    abstract::Heat shock protein 90 (Hsp90) is a fascinating target for cancer therapy due to its significant role in the crossroad of multiple signaling pathways associated with cell proliferation and regulation. Hsp90 inhibitors have the potential to be developed into anti-cancer drugs. Here, we identified nicotinic-mycoepoxydien...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmv034

    authors: Sun Y,Xiao S,Chen J,Wang M,Zheng Z,Song S,Zhang L

    更新日期:2015-06-01 00:00:00

  • Hyperhomocysteinemia-mediated DNA hypomethylation and its potential epigenetic role in rats.

    abstract::Hyperhomocysteinemia (HHcy), which is an independent risk factor for atherosclerosis, might cause dysregulation of gene expression, but the characteristics and key links involved in its pathogenic mechanisms are still poorly understood. The objective of the present study was to investigate the effect of HHcy on DNA me...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2007.00327.x

    authors: Jiang Y,Sun T,Xiong J,Cao J,Li G,Wang S

    更新日期:2007-09-01 00:00:00

  • A non-viral vector for potential DMD gene therapy study by targeting a minidystrophin-GFP fusion gene into the hrDNA locus.

    abstract::Gene therapy has emerged as a promising approach for the lethal disorder of Duchenne muscular dystrophy (DMD). Using a novel non-viral delivery system, the human ribosomal DNA (hrDNA) targeting vector, we targeted a minidystrophin-GFP fusion gene into the hrDNA locus of HT1080 cells with a high site-specific integrate...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmp080

    authors: Yang J,Liu X,Yu J,Sheng L,Shi Y,Li Z,Hu Y,Xue J,Wu L,Liang Y,Xia J,Liang D

    更新日期:2009-12-01 00:00:00

  • Effect of lentiviral vector-mediated KSR1 gene silencing on the proliferation of renal tubular epithelial cells and expression of inflammatory factors in a rat model of ischemia/reperfusion injury.

    abstract::Renal ischemia/reperfusion (I/R) is a common cause of acute renal failure in many clinical settings. Our study aimed to elucidate the role of lentiviral vector-mediated KSR1 gene silencing in inflammatory factor expression and proliferation of renal tubular epithelial cells (RTECs) in a rat model of I/R injury. Male S...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmy071

    authors: Bai Y,Han G,Guo K,Yu L,Du X,Xu Y

    更新日期:2018-08-01 00:00:00

  • HER4 promotes the growth and metastasis of osteosarcoma via the PI3K/AKT pathway.

    abstract::Osteosarcoma is the most common primary malignant bone tumor, which occurs in adolescents. As reported by our previous studies, HER4 indicates a poor prognosis of primary osteosarcoma. However, its mechanisms in the pathogenesis of osteosarcoma have not yet been studied. The purpose of this study was to investigate th...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmaa004

    authors: Li X,Huang Q,Wang S,Huang Z,Yu F,Lin J

    更新日期:2020-04-20 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

  • Regulation and functions of the Hippo pathway in stemness and differentiation.

    abstract::The Hippo pathway plays important roles in organ development, tissue regeneration, and human diseases, such as cancer. In the canonical Hippo pathway, the MST1/2-LATS1/2 kinase cascade phosphorylates and inhibits transcription coactivators Yes-associated protein and transcription coactivator with PDZ-binding motif and...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1093/abbs/gmaa048

    authors: Cao X,Wang C,Liu J,Zhao B

    更新日期:2020-07-10 00:00:00

  • Nucleic acid binding activity of pns6 encoded by genome segment 6 of rice ragged stunt oryzavirus.

    abstract::The ORF of genome segment 6 (S6) of rice ragged stunt oryzavirus (RRSV) Philippines isolate was cloned and sequenced based on the S6 sequence of the Thailand isolate. Pns6, the 71 kD product of S6 expressed in E. coli, was demonstrated to be a viral non-structural protein of RRSV by Western blotting. The gel mobility ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/36.7.457

    authors: Shao CG,Lü HJ,Wu JH,Gong ZX

    更新日期:2004-07-01 00:00:00

  • Chronic intermittent hypoxia promotes the development of experimental non-alcoholic steatohepatitis by modulating Treg/Th17 differentiation.

    abstract::The present study aims to characterize the effect of chronic intermittent hypoxia and HIF1α on the non-alcoholic steatohepatitis (NASH) process in mice, and to explore the role of the Treg/Th17 balance in the formation of NASH inflammation and fibrosis. To achieve this purpose, simple steatosis was induced in mice by ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmy131

    authors: Liu J,Li W,Zhu W,He W,Zhao H,Xiang Y,Liu C,Wu W

    更新日期:2018-12-01 00:00:00

  • Ricin-B-lectin enhances microsporidia Nosema bombycis infection in BmN cells from silkworm Bombyx mori.

    abstract::Nosema bombycis is an obligate intracellular parasitic fungus that utilizes a distinctive mechanism to infect Bombyx mori Spore germination can be used for host cell invasion; however, the detailed mechanism remains to be elucidated. The ricin-B-lectin (RBL) gene is significantly differentially regulated after N. bomb...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw093

    authors: Liu H,Li M,Cai S,He X,Shao Y,Lu X

    更新日期:2016-11-01 00:00:00

  • Emodin promotes the osteogenesis of MC3T3-E1 cells via BMP-9/Smad pathway and exerts a preventive effect in ovariectomized rats.

    abstract::Emodin, a natural anthraquinone extracted from the Chinese herbs rhubarb and giant knotweed rhizome, has been reported to enhance osteoblast differentiation. However, the mechanisms underlying its ability to regulate osteogenesis are unclear. The objective of this study was to determine the role of emodin in osteoblas...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmx087

    authors: Chen X,Zhang S,Chen X,Hu Y,Wu J,Chen S,Chang J,Wang G,Gao Y

    更新日期:2017-10-01 00:00:00

  • Cloning and overexpression of CYP6F1, a cytochrome P450 gene, from deltamethrin-resistant Culex pipiens pallens.

    abstract::CYP6F1 (GenBank/EMBL accession No. AY662654), a novel gene with a complete encoding sequence in the cytochrome P450 family 6, was cloned and sequenced from deltamethrin-resistant 4th instar larvae of Culex pipiens pallens. The cDNA sequence of CYP6F1 has an open reading frame of 1527 bp, which encodes a putative prote...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2005.00042.x

    authors: Gong MQ,Gu Y,Hu XB,Sun Y,Ma L,Li XL,Sun LX,Sun J,Qian J,Zhu CL

    更新日期:2005-05-01 00:00:00

  • Photothermal treatment with EGFRmAb-AuNPs induces apoptosis in hypopharyngeal carcinoma cells via PI3K/AKT/mTOR and DNA damage response pathways.

    abstract::Hypopharyngeal carcinoma (HC) is one of the most malignant tumors in the upper aerodigestive tract. Currently, there are no effective treatments for HC. Gold nanoparticles (AuNPs) are a promising tool that can be used for plasmonic photothermal therapy (PPTT), which refers to the use of electromagnetic radiation, most...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmy046

    authors: Zhang Y,Cong L,He J,Wang Y,Zou Y,Yang Z,Hu Y,Zhang S,He X

    更新日期:2018-06-01 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

  • The combined influence of surface modification, size distribution, and interaction time on the cytotoxicity of CdTe quantum dots in PANC-1 cells.

    abstract::Mercaptopropionic acid (MPA) and cysteamine (Cys) capped CdTe quantum dots (QDs) were successfully prepared and used to investigate the combined influence of surface modification, size distribution, and interaction time on their cytotoxicity in human pancreatic carcinoma (PANC-1) cells. Results indicated that the smal...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmr126

    authors: Chang S,Kang B,Liu X,Dai Y,Chen D

    更新日期:2012-03-01 00:00:00

  • PPAR-γ activation increases insulin secretion independent of CASK in INS-1 cells.

    abstract::Peroxisome proliferator-activated receptor-γ (PPAR-γ) is expressed in pancreatic β cells and is involved in insulin secretion. However, the precise mechanisms remain unclear. Calcium/calmodulin-dependent serine protein kinase (CASK), which plays a vital role in the anchoring of insulin granules on pancreatic β cell me...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmz052

    authors: Zhang K,Yuan Q,Xie J,Yuan L,Wang Y

    更新日期:2019-07-10 00:00:00

  • Edaravone injection reverses learning and memory deficits in a rat model of vascular dementia.

    abstract::Edaravone is a novel free radical scavenger that exerts neuroprotective effects by inhibiting endothelial injury and by ameliorating neuronal damage in brain ischemia. Recently, it was reported that edaravone could alleviate the pathology and cognitive deficits of Alzheimer's disease patients. However, its relevance t...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw116

    authors: Li X,Lu F,Li W,Qin L,Yao Y,Ge X,Yu Q,Liang X,Zhao D,Li X,Zhang J

    更新日期:2017-01-01 00:00:00

  • The Mycobacterium bovis BCG prime-Rv0577 DNA boost vaccination induces a durable Th1 immune response in mice.

    abstract::Tuberculosis remains a major global health problem and effective vaccines are urgently needed. In this study, we used the combined DNA- and protein-based vaccines of immunodominant antigen Rv0577 to boost BCG and evaluated their immunogenicity in BALB/c mice. Our data suggest that the booster vaccine may substantially...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw010

    authors: Gu D,Chen W,Mi Y,Gong X,Luo T,Bao L

    更新日期:2016-04-01 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