High-throughput proteomics: protein expression and purification in the postgenomic world.

Abstract:

:Proteomics has become a major focus as researchers attempt to understand the vast amount of genomic information. Protein complexity makes identifying and understanding gene function inherently difficult. The challenge of studying proteins in a global way is driving the development of new technologies for systematic and comprehensive analysis of protein structure and function. Protein expression and purification are key processes in these studies, but have typically only been applied on a case-by-case basis to proteins of interest. Researchers are addressing the challenge of parallel expression and purification of large numbers of gene products through the principles of high-throughput screening technologies commonly used in pharmaceutical development. Some of the issues relevant to these approaches are discussed here.

journal_name

Protein Expr Purif

authors

Lesley SA

doi

10.1006/prep.2001.1465

keywords:

subject

Has Abstract

pub_date

2001-07-01 00:00:00

pages

159-64

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(01)91465-2

journal_volume

22

pub_type

杂志文章,评审
  • Refolding and one-step purification of recombinant human ARA70 over-expressed in Escherichia coli.

    abstract::Androgen receptor (AR)-associated coregulator 70 (ARA70) is a cytoplasmic protein that has been characterized to have the ability to induce AR transcriptional activity in response to androgens and anti-androgens in prostate cancer cells. AR has been shown to have an important role in the progression of prostate cancer...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.11.002

    authors: Singh VK,Jia Z

    更新日期:2005-02-01 00:00:00

  • Efficient production of a correctly folded mouse α-defensin, cryptdin-4, by refolding during inclusion body solubilization.

    abstract::Mammalian α-defensins contribute to innate immunity by exerting antimicrobial activity against various pathogens. To perform structural and functional analysis of α-defensins, large amounts of α-defensins are essential. Although many expression systems for the production of recombinant α-defensins have been developed,...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.04.007

    authors: Tomisawa S,Sato Y,Kamiya M,Kumaki Y,Kikukawa T,Kawano K,Demura M,Nakamura K,Ayabe T,Aizawa T

    更新日期:2015-08-01 00:00:00

  • Effect of K225 residue to the catalytic efficiency of Kex2 protease.

    abstract::The gene encoding S. cerevisiae Kex2 protease derivative Kex2-667 (encoding the N-terminal 20th to 667th amino acid residues of Kex2 protease, containing the propeptide, catalytic domain, P domain and Ser/Thr enrichment region) and its 225th amino acid residue mutant K225L were overexpressed in Pichia pastoris. Protea...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2020.105725

    authors: Yang F,Liu L,Liu Y,Li S

    更新日期:2020-12-01 00:00:00

  • Molecular cloning, characterization, and heterologous expression of an acetyl-CoA acetyl transferase gene from Sanghuangporus baumii.

    abstract::Acetyl-CoA C-acetyltransferase synthase gene (AACT) cDNA, DNA and promoter were cloned from Sanghuangporus baumii. The gene ORF (1260 bp) encoded 419 amino acids. The AACT DNA includes five exons (1-84 bp, 140-513 bp, 570-1027 bp, 1090-1282 bp, 1344-1494 bp) and four introns (85-139 bp, 514-569 bp, 1028-1089 bp, 1283-...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2020.105592

    authors: Wang X,Wang S,Xu X,Sun J,Ma Y,Liu Z,Sun T,Zou L

    更新日期:2020-06-01 00:00:00

  • Overproduction, purification, and characterization of recombinant bifunctional threonine-sensitive aspartate kinase-homoserine dehydrogenase from Arabidopsis thaliana.

    abstract::In plant, the first and the third steps of the synthesis of methionine and threonine are catalyzed by a bifunctional enzyme, aspartate kinase-homoserine dehydrogenase (AK-HSDH). In this study, we report the first purification and characterization of a highly active threonine-sensitive AK-HSDH from plants (Arabidopsis ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2001.1539

    authors: Paris S,Wessel PM,Dumas R

    更新日期:2002-02-01 00:00:00

  • Expression and bioactivity analysis of Staphylococcal enterotoxin M and N.

    abstract::Staphylococcal enterotoxins (SEs) are powerful superantigens that stimulate non-specific T-cell proliferation produced by Staphylococcus aureus and draw considerable attention as ideal drugs for cancer therapy. The filtrate of S. aureus culture has been used as ampul named Staphylococcal enterotoxin C injection in cli...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2007.08.005

    authors: Pan YQ,Ding D,Li DX,Chen SQ

    更新日期:2007-12-01 00:00:00

  • Remedial strategies in structural proteomics: expression, purification, and crystallization of the Vav1/Rac1 complex.

    abstract::The signal transduction pathway involving the Vav1 guanine nucleotide exchange factor (GEF) and the Rac1 GTPase plays several key roles in the immune response mediated by the T cell receptor. Vav1 is also a unique member of the GEF family in that it contains a cysteine-rich domain (CRD) that is critical for Rac1 bindi...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2006.10.027

    authors: Brooun A,Foster SA,Chrencik JE,Chien EY,Kolatkar AR,Streiff M,Ramage P,Widmer H,Weckbecker G,Kuhn P

    更新日期:2007-05-01 00:00:00

  • Enhanced soluble expression of recombinant Flavobacterium heparinum heparinase I in Escherichia coli by fusing it with various soluble partners.

    abstract::Heparinase I (HepA) was originally isolated from Flavobacterium heparinum (F. heparinum) and specifically cleaves heparin/heparan sulfate in a site-dependent manner, showing great promise for producing low molecular weight heparin (LMWH). However, expressing recombinant HepA is extremely difficult in Escherichia coli ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2012.03.016

    authors: Huang J,Cao L,Guo W,Yuan R,Jia Z,Huang K

    更新日期:2012-06-01 00:00:00

  • Cloning, expression, purification, and characterization of a glutamate-specific endopeptidase from Bacillus licheniformis.

    abstract::A gene encoding a glutamate-specific endopeptidase (GSE) from Bacillus licheniformis (BL) has been cloned in Escherichia coli cells. The recombinant protein was expressed as cytoplasmic insoluble inclusion bodies. Immobilized metal affinity chromatography was employed to purify the protein, and then a 27-kDa GSE inter...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2011.12.001

    authors: Ye W,Liu J,Wang H,Wang J,Wang X

    更新日期:2012-03-01 00:00:00

  • Purification and characterization of the L-Ara4N transferase protein ArnT from Salmonella typhimurium.

    abstract::The covalent addition of 4-amino-4-deoxy-L-arabinose (L-Ara4N) groups to lipid A, which resides in the outer membranes of bacteria such as Salmonella typhimurium and Escherichia coli, is the final step in the polymyxin-resistance pathway in these organisms. This modification is catalyzed by the inner membrane protein ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2005.08.028

    authors: Bretscher LE,Morrell MT,Funk AL,Klug CS

    更新日期:2006-03-01 00:00:00

  • Expression, purification, and characterization of the recombinant NAD-malic enzyme from Ascaris suum.

    abstract::The cDNA encoding the 65-kDa subunit of malic enzyme from Ascaris suum was cloned into the bacterial expression vector pKK223-3 and overproduced in Escherichia coli. A protein with a subunit molecular mass of 65,000 was expressed at a level of up to 3% of the total soluble protein in JM109, as judged by SDS-PAGE. The ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0705

    authors: Chooback L,Karsten WE,Kulkarni G,Nalabolu SR,Harris BG,Cook PF

    更新日期:1997-06-01 00:00:00

  • Heterologous expression and purification of plantaricin NC8, a two-peptide bacteriocin against Salmonella spp. from Lactobacillus plantarum ZJ316.

    abstract::Bacteriocin, which is produced by lactic acid bacteria (LAB), has the potential to act as natural preservatives in the food industry. To develop strategies to overproduce such peptides, plantaricin NC8, a class IIb LAB bacteriocin that consists of two peptides, PLNC8α and PLNC8β, was successfully heterologously expres...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2016.06.013

    authors: Jiang H,Li P,Gu Q

    更新日期:2016-11-01 00:00:00

  • Isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow's milk using gel filtration and anion-exchange chromatography including evaluation of their antigenicity.

    abstract::The aim of this study was to introduce a simple, reproducible, and less expensive method for isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow's milk while retaining their antigenicity. Whey (lactoserum) was obtained by isolating casein from defatted milk using hydrochloric acid. Gl...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(03)00015-9

    authors: Neyestani TR,Djalali M,Pezeshki M

    更新日期:2003-06-01 00:00:00

  • Immediate-early baculovirus vectors for foreign gene expression in transformed or infected insect cells.

    abstract::Baculovirus expression vectors are used routinely for foreign gene expression and are under intense development as improved biological pesticides. Conventional baculovirus expression vectors are recombinant viruses that can express a foreign gene in insect cells under the control of the polyhedrin promoter, which prov...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0092

    authors: Jarvis DL,Weinkauf C,Guarino LA

    更新日期:1996-09-01 00:00:00

  • Improving storage stability of recombinant organophosphorus hydrolase.

    abstract::Organophosphorus hydrolase (OPH) is a ∼38kDa enzyme encoded by opd gene of Flavobacterium sp. The enzyme can hydrolyze and inactivate variety of organophosphate (OP)-compounds, including chemical warfare nerve agents. Thus, OPH is a strong candidate for the development of therapeutic intervention against OP-poisoning ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.01.012

    authors: Satvik Iyengar AR,Tripathy RK,Bajaj P,Pande AH

    更新日期:2015-07-01 00:00:00

  • Expression and characterization of Pen b 26 allergen of Penicillium brevicompactum in Escherichia coli.

    abstract::Pen b 26 is one of the allergens produced by Penicillium brevicompactum which is one of the most prevalent in door airborne fungi and a major source of respiratory problems, including asthma. Pen b 26 wa scloned and expressed as an N-terminal as well as a C-terminal His6 tagged fusion protein in Escherichia coli. This...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2009.01.004

    authors: Sevinc MS,Kumar V,Abebe M,Mohottalage S,Kumarathasan P,Vincent R,Vijay HM

    更新日期:2009-05-01 00:00:00

  • Transmembrane-sequence-dependent overexpression and secretion of glycoproteins in Saccharomyces cerevisiae.

    abstract::Protein expression using the secretory pathway in Saccharomyces cerevisiae can lead to high amounts of overexpressed and secreted proteins in culture supernatants in a short period of time. These post-translational modified expression products can be purified up to >90% in a single step. The overexpression and secreti...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2000.1337

    authors: Schuster M,Wasserbauer E,Aversa G,Jungbauer A

    更新日期:2001-02-01 00:00:00

  • Optimization of expression and purification of two biologically active chimeric fusion proteins that consist of human interleukin-13 and Pseudomonas exotoxin in Escherichia coli.

    abstract::We have previously reported that a variety of solid human tumor cell lines express a large number of receptors for interleukin-13 (IL-13). These receptors could be targeted with a chimeric fusion protein consisting of human IL-13 and a truncated form of Pseudomonas exotoxin (PE). We describe here optimization of criti...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.10.012

    authors: Joshi BH,Puri RK

    更新日期:2005-02-01 00:00:00

  • Improvements of a high-throughput protein purification process using a calcium-dependent setup.

    abstract::The Human Secretome Project aims to produce and purify all human secreted proteins as full-length. In order to enable this, a robust, gentle and effective purification process is needed, where multiple proteins can be purified in parallel. For this reason, a purification system based on a Protein C-tag and the HPC4 an...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2020.105698

    authors: Kanje S,Enstedt H,Dannemeyer M,Uhlén M,Hober S,Tegel H

    更新日期:2020-11-01 00:00:00

  • Expression of Pisum sativum SAD polypeptides in production hosts and in planta: tetrameric organization of the protein.

    abstract::In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.09.004

    authors: Scherbak N,Brosché M,Ala-Häivälä A,Strid H,Ohrfelt A,Nilsson F,Strid A

    更新日期:2009-01-01 00:00:00

  • Expression in E. coli and purification of the nucleoside diphosphate kinase b from Leishmania major.

    abstract::Leishmaniasis is considered by the World Health Organization to be the second most important disease caused by a protozoan parasite. Biochemical and molecular biology studies can help in the understanding of the biology of the Leishmania parasite. All protozoan parasites, including Leishmania, are unable to synthesize...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2006.05.010

    authors: de Oliveira AH,Ruiz JC,Cruz AK,Greene LJ,Rosa JC,Ward RJ

    更新日期:2006-10-01 00:00:00

  • Expression, refolding and purification of Ov-GRN-1, a granulin-like growth factor from the carcinogenic liver fluke, that causes proliferation of mammalian host cells.

    abstract::Granulins (GRNs) are potent growth factors that are upregulated in many aggressive cancers from a wide range of organs. GRNs form tight, disulphide bonded, beta hairpin stacks, making them difficult to express in recombinant form. We recently described Ov-GRN-1, a GRN family member secreted by the carcinogenic liver f...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2011.06.018

    authors: Smout MJ,Mulvenna JP,Jones MK,Loukas A

    更新日期:2011-10-01 00:00:00

  • High-level synthesis of recombinant murine endostatin in Chinese hamster ovary cells.

    abstract::Endostatin, a carboxy-terminal fragment of collagen XVIII, has been shown to act as an anti-angiogenic agent that specifically inhibits proliferation of endothelial cells and growth of various primary tumors. Here, we describe the expression by Chinese hamster ovary (CHO) cells of murine endostatin and of a tagged-fus...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.01.003

    authors: Chura-Chambi RM,Tornieri PH,Spencer PJ,Nascimento PA,Mathor MB,Morganti L

    更新日期:2004-05-01 00:00:00

  • Expression of the Trichoderma reesei tyrosinase 2 in Pichia pastoris: isotopic labeling and physicochemical characterization.

    abstract::Trichoderma reesei tyrosinase TYR2 has been demonstrated to be able to oxidize various phenolic compounds and also peptide and protein bound tyrosine, and thus is of great interest for different biotechnological applications. In order to understand the reaction mechanism of the enzyme it would be essential to solve it...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2007.04.014

    authors: Westerholm-Parvinen A,Selinheimo E,Boer H,Kalkkinen N,Mattinen M,Saloheimo M

    更新日期:2007-09-01 00:00:00

  • Functional expression of N-terminally tagged membrane bound cytochrome P450.

    abstract::The introduction of an affinity tag offers an attractive approach to isolation of membrane proteins. The type of affinity tag and its positioning in the protein is determined by the desired subsequent experimental uses of the isolated protein. To minimize the risk of interference, membrane proteins may preferentially ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2009.06.005

    authors: Hamann T,Laursen T,Møller BL

    更新日期:2009-11-01 00:00:00

  • Soluble expression in Escherichia coli, one-step purification, and characterization of Chinese hamster dihydrofolate reductase.

    abstract::Chinese hamster dihydrofolate reductase (ch-DHFR) was overexpressed in Escherichia coli DH5 alpha under the transcriptional control of PRPL promoters regulated by temperature-sensitive repressors. The desired recombinant product is soluble and constitutes about 30% of the total soluble proteins of the bacterial cell. ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0676

    authors: Fan YX,Wang B,Jing GZ

    更新日期:1997-03-01 00:00:00

  • Overexpression and purification of the Escherichia coli inner membrane enzyme acyl-acyl carrier protein synthase in an active form.

    abstract::Acyl-acyl carrier protein synthase (Aas) is widely used to synthesize thioester adducts of fatty acids between 8 and 18 carbons in length enzymatically to the phosphopantetheine group of acyl carrier protein. The enzyme is an 80.6-kDa inner membrane protein that functions in vivo as a 2-acylglycerophosphoethanolamine ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2000.1206

    authors: Shanklin J

    更新日期:2000-04-01 00:00:00

  • Production and purification of a recombinant human hsp60 epitope using the cellulose-binding domain in Escherichia coli.

    abstract::The heat shock protein hsp60 plays a functional role in insulin-dependent diabetes mellitus. The hsp60 epitope p277 (aa 437-aa 460) is effective in vaccinating mice against diabetes. A synthetic peptide gene (p277) that encodes the human hsp60 epitope was cloned to the 3' end of the cellulose-binding domain gene (cbd)...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1998.0929

    authors: Shpigel E,Elias D,Cohen IR,Shoseyov O

    更新日期:1998-11-01 00:00:00

  • Formation of well-defined soluble aggregates upon fusion to MBP is a generic property of E6 proteins from various human papillomavirus species.

    abstract::Protein aggregation is a main barrier hindering structural and functional studies of a number of interesting biological targets. The E6 oncoprotein of Human Papillomavirus strain 16 (E6(16)) is difficult to express under a native soluble form in bacteria. Produced as an unfused sequence, it forms inclusion bodies. Fus...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2006.07.029

    authors: Zanier K,Nominé Y,Charbonnier S,Ruhlmann C,Schultz P,Schweizer J,Travé G

    更新日期:2007-01-01 00:00:00

  • Characterization of ubiquitin C-terminal hydrolase 1 (YUH1) from Saccharomyces cerevisiae expressed in recombinant Escherichia coli.

    abstract::The YUH1 gene coding for ubiquitin C-terminal hydrolase 1, a deubiquitinating enzyme, was cloned from the Saccharomyces cerevisiae genomic DNA and expressed in Escherichia coli. YUH1 was fused with the 6 histidine tag at the N-terminus (H6YUH1) or C-terminus (YUH1H6) and purified by an immobilized metal affinity chrom...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2007.07.005

    authors: Yu HA,Kim SG,Kim EJ,Lee WJ,Kim DO,Park K,Park YC,Seo JH

    更新日期:2007-11-01 00:00:00