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-expressed as a fusion to the N-terminus of a self-cleavable affinity tag. After the PACAP-intein-CBD fusion protein was purified by chitin-affinity chromatography, the self-cleavage activity of the intein was induced by DTT and the rhPACAP was released from the chitin-bound intein tag. The activity of the rhPACAP to stimulate cyclic AMP accumulation was detected using the human pancreas carcinoma cells SW1990. Twenty-two milligrams of rhPACAP with the purity over 98% was obtained by single column purification from 1 liter of induced culture. The preliminary biological assay indicated that the rhPACAP, which has an extra Met at its N-terminus compared with the native human PACAP, had the similar activity of stimulating cAMP accumulation with the standard PACAP38 in the SW1990 cells. A new efficient production procedure of the active recombinant human PACAP was established.

authors

Yu RJ,Hong A,Dai Y,Gao Y

doi

10.1093/abbs/36.11.759

subject

Has Abstract

pub_date

2004-11-01 00:00:00

pages

759-66

issue

11

eissn

1672-9145

issn

1745-7270

journal_volume

36

pub_type

杂志文章
  • 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

  • The effects and regulatory mechanism of RIP3 on RGC-5 necroptosis following elevated hydrostatic pressure.

    abstract::Necroptosis is a type of regulated cell death that has been implicated in various diseases. Receptor-interacting protein 3 (RIP3), a member of the RIP family, is an important mediator of the necroptotic pathway. Cleavage of RIP3 at Asp328 by caspase-8 abolishes the kinase activity of RIP3, which is critical for necrop...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw130

    authors: Shang L,Ding W,Li N,Liao L,Chen D,Huang J,Xiong K

    更新日期:2017-02-06 00:00:00

  • Role of chaperone-mediated autophagy in degrading Huntington's disease-associated huntingtin protein.

    abstract::Mutant N-terminal huntingtin (Htt) protein resulting from Huntington's disease (HD) with expanded polyglutamine accumulates and forms aggregates in vulnerable neurons. Both ubiquitin proteasomal and autophagic pathways contribute to the degradation of mutant Htt. Here, we focus on the involvement of chaperone-mediated...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1093/abbs/gmt133

    authors: Qi L,Zhang XD

    更新日期:2014-02-01 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

  • Ultrasound/microbubble enhances foreign gene expression in ECV304 cells and murine myocardium.

    abstract::Although viral vectors are efficient systems to transfer foreign genes into cells or target tissues, safety issues remain in relation to human gene therapy. Microbubbles currently used as ultrasound contrast agents have been applied in transfection of genes. This study was designed to test the transfection efficiency ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/36.12.824

    authors: Guo DP,Li XY,Sun P,Wang ZG,Chen XY,Chen Q,Fan LM,Zhang B,Shao LZ,Li XR

    更新日期:2004-12-01 00:00:00

  • Biological characteristics of dengue virus and potential targets for drug design.

    abstract::Dengue infection is a major cause of morbidity in tropical and subtropical regions, bringing nearly 40% of the world population at risk and causing more than 20,000 deaths per year. But there is neither a vaccine for dengue disease nor antiviral drugs to treat the infection. In recent years, dengue infection has been ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1111/j.1745-7270.2008.00382.x

    authors: Qi RF,Zhang L,Chi CW

    更新日期:2008-02-01 00:00:00

  • Computational analysis of microRNA function in heart development.

    abstract::Emerging evidence suggests that specific spatio-temporal microRNA (miRNA) expression is required for heart development. In recent years, hundreds of miRNAs have been discovered. In contrast, functional annotations are available only for a very small fraction of these regulatory molecules. In order to provide a global ...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmq072

    authors: Liu G,Ding M,Chen J,Huang J,Wang H,Jing Q,Shen B

    更新日期:2010-09-01 00:00:00

  • Fusion protein of interleukin 4 and diphtherial toxin with high cytotoxicity to cancer cells.

    abstract::Receptor of human interleukin 4 (hIL4R) has been found to be present on many types of cancer, so it may be a good target for cancer therapy. Here, fusion toxin gene DT4H has been constructed by fusing DNA sequence encoding the first 389 amino acids of diphtherial toxin (DT), which can not bind its own receptor, to hum...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/36.6.437

    authors: Zhang YJ,Hu XB,Li SX,Tian LP,Yang SL,Gong Y

    更新日期:2004-06-01 00:00:00

  • MicroRNA expression profile of gastric cancer stem cells in the MKN-45 cancer cell line.

    abstract::MicroRNAs (miRNAs) play important roles in post-transcriptional gene silencing of target messenger RNAs, which are involved in virtually all biological processes. Previously, we have demonstrated that spheroid body-forming cells from the MKN-45 cancer cell line possessed gastric cancer stem cell (CSC) properties. In t...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmt135

    authors: Liu J,Ma L,Wang Z,Wang L,Liu C,Chen R,Zhang J

    更新日期:2014-02-01 00:00:00

  • Establishment and utilization of a tetracycline-controlled inducible RNA interfering system to repress gene expression in chronic myelogenous leukemia cells.

    abstract::RNA interference (RNAi), a posttranscriptional gene silencing process mediated by small double-stranded RNA specifically complementary to the targeted transcript, has been used extensively in the development of novel therapeutic approaches against various human diseases including chronic myelogenous leukemia (CML). He...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2005.00112.x

    authors: Yang F,Zhang Y,Cao YL,Wang SH,Liu L

    更新日期:2005-12-01 00:00:00

  • Knockdown of STAT3 by shRNA inhibits the growth of CAOV3 ovarian cancer cell line in vitro and in vivo.

    abstract::Constitutively activated signal transducer and activator of transcription 3 (STAT3) plays an important role in the formation of many tumors including ovarian cancer. In this study, RNA interference specific to STAT3 was employed to study its effects on the inhibition of STAT3 signaling and on the growth of ovarian can...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2008.00424.x

    authors: Huang F,Tong X,Fu L,Zhang R

    更新日期:2008-06-01 00:00:00

  • New hope in the horizon: cancer stem cells.

    abstract::The major goal of researchers and oncologists is to develop promising ground for novel therapeutic strategies to prevent recurrence or relapse of cancer. Recent evidences suggest that a subset of cells called cancer stem cells (CSCs) are present within the tumor mass which possess tumorigenic capacity and may be respo...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1093/abbs/gmq013

    authors: Bhattacharyya S,Khanduja KL

    更新日期:2010-04-01 00:00:00

  • Effect of exogenous ghrelin on cell differentiation antigen 40 expression in endothelial cells.

    abstract::Ghrelin is a brain-gut peptide that serves as a natural ligand for growth hormone secretagogue receptor (GHSR). It also exists abundantly in the cardiovascular system. In order to evaluate the possible role of ghrelin in the development of atherosclerosis, the effect of ghrelin on the expression of cell differentiatio...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2007.00365.x

    authors: Zhang M,Yuan F,Chen H,Qiu X,Fang W

    更新日期:2007-12-01 00:00:00

  • N6-methyladenosine methyltransferase plays a role in hypoxic preconditioning partially through the interaction with lncRNA H19.

    abstract::N6-methyladenosine (m6A), a methylation in the N6 position of adenosine especially in the mRNA, exerts diverse physiological and pathological functions. However, the precise role of m6A methylation in hypoxic preconditioning (HPC) is still unknown. Here, we observed that HPC treatment protected H9c2 cells against H2O2...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmaa130

    authors: Su Y,Xu R,Zhang R,Qu Y,Zuo W,Ji Z,Geng H,Pan M,Ma G

    更新日期:2020-12-29 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

  • 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

  • Biological activities of purified harpin(Xoo) and harpin(Xoo) detection in transgenic plants using its polyclonal antibody.

    abstract::Many harpins have been found in plant pathogen bacteria that can elicit disease and insect resistance in plants, and promote plant growth. In this work, we overexpressed and purified Xanthomonas oryzae pv. oryzae harpin, harpin(Xoo), in Escherichia coli BL21/pGEX-hpa1. Harpin(Xoo) was fused to the C-terminus of glutat...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2005.00096.x

    authors: Li M,Shao M,Lu XZ,Wang JS

    更新日期:2005-10-01 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 subunit vaccine based on rH-NS induces protection against Mycobacterium tuberculosis infection by inducing the Th1 immune response and activating macrophages.

    abstract::Mycobacterium tuberculosis (Mtb) is a Gram-positive pathogen which causes tuberculosis in both animals and humans. All tested rH-NS formulations induced a specific Th1 response, as indicated by increased production of interferon γ (IFN-γ) and interleukin 2 (IL-2) by lymphocytes in the spleen of mice which were immuniz...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw078

    authors: Liu Y,Chen S,Pan B,Guan Z,Yang Z,Duan L,Cai H

    更新日期:2016-10-01 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

  • Predicting polymerase II core promoters by cooperating transcription factor binding sites in eukaryotic genes.

    abstract::Several discriminate functions for predicting core promoters that based on the potential cooperation between transcription factor binding sites (TFBSs) are discussed. It is demonstrated that the promoter predicting accuracy is improved when the cooperation among TFBSs is taken into consideration. The core promoter reg...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/36.4.250

    authors: Ma XT,Qian MP,Tang HX

    更新日期:2004-04-01 00:00:00

  • The regulation and function of YAP transcription co-activator.

    abstract::The Hippo pathway was initially identified in Drosophila by genetic mosaic screens for tumor suppressor genes. Researches indicated that the Hippo pathway is a key regulator of organ size and is conserved during evolution. Furthermore, studies of mouse models and clinical samples demonstrated the importance of Hippo p...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章,评审

    doi:10.1093/abbs/gmu110

    authors: Zhu C,Li L,Zhao B

    更新日期:2015-01-01 00:00:00

  • Adenovirus-mediated co-expression of ING4 and PTEN cooperatively enhances their antitumor activity in human hepatocellular carcinoma cells.

    abstract::Both inhibitor of growth 4 (ING4) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) are well known as tumor suppressors that are closely related to tumor occurrence and progression. It was reported that ING4 and PTEN showed synergistic antitumor activities in nasopharyngeal carcinoma cells. The two tu...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmw062

    authors: Rakshit N,Yang S,Zhou W,Xu Y,Deng C,Yang J,Yu H,Wei W

    更新日期:2016-08-01 00:00:00

  • Oleanolic acid induces apoptosis in human leukemia cells through caspase activation and poly(ADP-ribose) polymerase cleavage.

    abstract::It has been shown that Fructus Ligustri Lucidi (FLL), a promising traditional Chinese medicine, can inhibit the growth of tumors. However, the effective component and molecular mechanism of FLL act to inhibit tumor proliferation are unclear. In this study, we demonstrated that oleanolic acid (OA), a principal chemical...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2007.00335.x

    authors: Zhang P,Li H,Chen D,Ni J,Kang Y,Wang S

    更新日期:2007-10-01 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

  • 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

  • Crystal structures of human dipeptidyl peptidase IV in its apo and diprotin B-complexed forms.

    abstract::Dipeptidyl peptidase IV (DPPIV), which belongs to the prolyl oligopeptidase family of serine proteases, is known to have a variety of regulatory biological functions and has been shown to be implicated in type 2 diabetes. It is therefore important to develop selective human DPPIV (hDPPIV) inhibitors. In this study, we...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2007.00289.x

    authors: Hiramatsu H,Kyono K,Yamamoto A,Saeki K,Shima H,Sugiyama S,Inaka K,Shimizu R

    更新日期:2007-05-01 00:00:00

  • Establishment and application of a TaqMan real-time quantitative reverse transcription-polymerase chain reaction assay for rubella virus RNA.

    abstract::The aim of this study was to establish and apply a real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) for rubella virus (RV) RNA. First, the primer and TaqMan probe concentrations, as well as reaction temperatures were optimized to establish an efficient real-time quantitative RT-PCR assay...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1111/j.1745-7270.2006.00213.x

    authors: Zhao LH,Ma YY,Wang H,Zhao SP,Zhao WM,Li H,Wang LY

    更新日期:2006-10-01 00:00:00

  • MiRNA-218, a new regulator of HMGB1, suppresses cell migration and invasion in non-small cell lung cancer.

    abstract::MicroRNAs (miRNAs) function as negative regulators of gene expression involved in cancer metastasis. The aim of this study is to investigate the potential roles of miR-218 in non-small cell lung cancer and validate its regulation mechanism. Functional studies showed that miR-218 overexpression inhibited cell migration...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmt109

    authors: Zhang C,Ge S,Hu C,Yang N,Zhang J

    更新日期:2013-12-01 00:00:00

  • The inhibitory effect of microRNA-1827 on anoikis resistance in lung adenocarcinoma A549 cells via targeting caveolin-1.

    abstract::Anoikis resistance is a critical process for cancer cell metastasis in non-small cell lung cancer (NSCLC), and microRNA-1827 (miR-1827) is closely correlated with NSCLC metastasis. In this study, we aimed to evaluate the role of miR-1827 in regulating the anoikis resistance of NSCLC. The results showed that miR-1827 l...

    journal_title:Acta biochimica et biophysica Sinica

    pub_type: 杂志文章

    doi:10.1093/abbs/gmaa102

    authors: Guo X,Wang Z,Sun Q,Sun C,Hua H,Huang Q

    更新日期:2020-10-19 00:00:00