Abstract:
:A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domain is truncated at the end of a predicted helix and prior to a Q-linker. The domain was found to be very soluble and stable so that it could be purified to homogeneity and crystallized. This construct has deadenylylation activity that is independent of the low nitrogen status indicator PII-UMP. The crystals belong to space group P3(1)21 or its enantiomorph P3(2)21 with a=b=116.6 A and c=67.6 A.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Xu Y,Wen D,Clancy P,Carr PD,Ollis DL,Vasudevan SGdoi
10.1016/j.pep.2003.11.001keywords:
subject
Has Abstractpub_date
2004-03-01 00:00:00pages
142-6issue
1eissn
1046-5928issn
1096-0279pii
S1046592803003668journal_volume
34pub_type
杂志文章abstract::In this report, we describe an optimized system for the efficient overexpression, purification, and refolding of secreted bacterial proteins. Candidate secreted proteins were produced recombinantly in Escherichia coli as Tobacco Etch Virus protease-cleavable hexahistidine-c-myc eptiope fusion proteins. Without regard ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.09.003
更新日期:2006-03-01 00:00:00
abstract::Procedures are described for the preparation of highly purified thymidylate synthases from Escherichia coli and Bacillus subtilis. The yields in each case are quite high with about 350 mg of pure protein obtained from 1 liter of cells. Basically all that is required to obtain pure enzyme is an induction step from a hi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1245
更新日期:2000-07-01 00:00:00
abstract::Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.06.011
更新日期:2006-11-01 00:00:00
abstract::Deformylation of the initiator N-formylmethionine does not always proceed to completion for proteins overexpressed in Escherichia coli. To overcome this limitation, the def gene encoding the Escherichia coli peptide deformylase was cloned into the plysS plasmid under the tetracycline (Tc) promoter control. The efficie...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.03.007
更新日期:2004-07-01 00:00:00
abstract::The TCL1 gene, which is located on chromosome 14, plays a major role in human hematopoietic malignancies and encodes a 14-kDa protein whose function has not been determined. This gene is expressed in pre-B cells, in immature thymocytes, and, at low levels, in activated T cells but not in peripheral mature B cells and ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1186
更新日期:2000-04-01 00:00:00
abstract::Human Fas ligand is a medically important membrane glycoprotein that induces the apoptosis of harmful cells. A new secretory expression and purification method was devised for the production of a large amount of recombinant human Fas ligand extracellular domain (hFasLECD) by Pichia pastoris. The expression plasmid con...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.08.006
更新日期:2006-12-01 00:00:00
abstract::Methionine aminopeptidases (MetAPs), ubiquitous enzymes that play an important role in nascent protein maturation, have been recognized as attractive targets for the development of drugs against pathogenic protozoa including Plasmodium spp. Here, we characterized partial biochemical properties of a type I MetAP of Pla...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.01.003
更新日期:2015-04-01 00:00:00
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
更新日期:2005-02-01 00:00:00
abstract::L1 is a human cell adhesion glycoprotein involved in the development of the central nervous system that comprises six immunoglobulin-like domains (Ig1-Ig6), five fibronectin-type III (FN1-FN5) domains, a single transmembrane region and a cytoplasmic domain. It contains 20 potential N-glycosylation sites and is heavily...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.10.008
更新日期:2007-03-01 00:00:00
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
更新日期:2007-05-01 00:00:00
abstract::FcgammaRs are involved in regulating a multitude of innate and adaptive immune responses, which makes them attractive targets for the development of novel immunotherapeutic approaches. In this report, we describe a simple method for the production of a large quantity of recombinant porcine FcgammaRII. The extracellula...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.06.020
更新日期:2009-11-01 00:00:00
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
更新日期:2009-05-01 00:00:00
abstract::Proteins are essential throughout the biological and biomedical sciences and the purification strategies of proteins of interest have advanced over centuries. Elastin-like polypeptides (ELPs) are compound polymers that have recently been highlighted for their sharp and reversible phase transition property when heated ...
journal_title:Protein expression and purification
pub_type: 杂志文章,评审
doi:10.1016/j.pep.2018.09.006
更新日期:2019-01-01 00:00:00
abstract::We present a straightforward, versatile method for expressing and purifying β-amyloid (Aβ40) and transmembrane peptides derived from β-amyloid precursor protein (Aβ55). In principle, these methods should be applicable to other types of strongly aggregating peptides. We start with a DNA plasmid encoding a HexaHis tag w...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.04.006
更新日期:2019-10-01 00:00:00
abstract::Insulin-like growth factor binding proteins (IGFBPs) display many functions in humans including regulation of the insulin-like growth factor (IGF) signaling pathway. The various roles of human IGFBPs make them attractive protein candidates in drug discovery. Structural and functional knowledge on human proteins with t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.10.017
更新日期:2015-04-01 00:00:00
abstract::To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to create a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-length clone by exonuclease III. When a construct is equipped ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.12.013
更新日期:2005-05-01 00:00:00
abstract::Anthrax toxin consists of three separate proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA binds to the receptor on mammalian cells and facilitates translocation of EF or LF into the cytosol. PA is the primary component of several anthrax vaccines. In this study we expressed and purified...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0005
更新日期:1996-02-01 00:00:00
abstract::Pseudomonas fluorescens is a robust protein expression system that is very well suited for high throughput protein expression for structural genomics studies. Since NMR spectroscopy and X-ray crystallography are both used by various investigators in structure elucidation studies, the availability of target proteins la...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.12.012
更新日期:2009-05-01 00:00:00
abstract::A long-lasting recombinant human albumin-linker-erythropoietin (EPO) is a human albumin gene fused to the N-terminal of EPO with a (GGSGG)(n)-repeated linker inserted between albumin and EPO. Albumin-EPO fusion genes were co-transfected with the dhfr gene. Albumin-EPO fusion protein has three kinds of sub-types (IALE,...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.07.003
更新日期:2009-12-01 00:00:00
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
更新日期:2015-08-01 00:00:00
abstract::We previously reported the expression and purification of recombinant p68 RNA helicase in a bacterial expression system. The recombinant p68 is an RNA-dependent ATPase and ATP-dependent RNA helicase. In the process of characterizing the ATPase and RNA unwinding activities of the recombinant p68, we observed that the b...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.02.004
更新日期:2004-06-01 00:00:00
abstract::The erm proteins confer resistance to the MLS (macrolide-lincosamide-streptogramin B) antibiotics in various microorganisms, including pathogens, through dimethylation of a single adenine residue (A2085: Bacillus subtilis coordinate) of the 23S rRNA to reduce the affinity of antibiotics, thereby enabling the cells to ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2002.1621
更新日期:2002-06-01 00:00:00
abstract::The zoonotic Rift Valley fever virus affects livestock and humans in Africa and on the Arabian Peninsula. The economic impact of this pathogen due to livestock losses, as well as its relevance to public health, underscores the importance of developing effective and easily distributed vaccines. Vaccines that can be del...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.07.003
更新日期:2016-11-01 00:00:00
abstract::We describe a single method to purify milligram amounts of authentic wild-type and mutant HIV-1 and HIV-2 Tat proteins overexpressed in Escherichia coli. This method takes advantage of the highly basic, positively charged RNA binding domain present in both HIV-1 and HIV-2 Tat, which also facilitated purification of HI...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0096
更新日期:1996-09-01 00:00:00
abstract::Cyclohexanone monooxygenase (CMO) is a soluble flavoenzyme originally isolated from Acinetobacter spp. which carries out Baeyer-Villiger reactions with cyclic ketone substrates. In the present study we cloned the Acinetobacter CMO gene and modified it for facile purification from heterologous expression systems by inc...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1340
更新日期:2001-02-01 00:00:00
abstract::Ciliary neurotrophic factor (CNTF) is characterized as a neuropoietic cytokine for a broad spectrum of neurons, leading to its evaluation in humans suffering from neurodegenerative diseases. Due to its wide range of biological applications, high yield production of soluble biologically active recombinant human CNTF (r...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.01.008
更新日期:2014-04-01 00:00:00
abstract::Campylobacteriosis is a disease in humans caused by the infection from Campylobacter spp. Human cases are mainly due to Campylobacter jejuni, although C. coli can cause gastroenteritis in humans as well. The bacteria are commensal in chicken tract and can be contaminated into chicken products during processing. Obviou...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2021.105818
更新日期:2021-04-01 00:00:00
abstract::MMP1 is an essential enzyme for tissue remodeling both in normal and pathological states. We report a method of purifying activated human MMP1 in E. coli without using urea or 4-Aminophenylmercuric acetate (APMA). Instead, a non-ionic detergent, Triton X-100, was used in the lysis buffer to solubilize MMP1 followed by...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.04.001
更新日期:2018-08-01 00:00:00
abstract::The fusion protein consisting of human Fas receptor extracellular domain and human IgG1 heavy chain Fc domain (hFasRECD-Fc) is a medically important protein that potentially has therapeutic uses. The fusion gene composed of a synthetic human Fas receptor extracellular domain gene and the cDNA encoding human IgG1 heavy...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.05.007
更新日期:2010-10-01 00:00:00
abstract::Previously, the lysozyme gene of the Klebsiella phage K11 was partially sequenced in our lab. Using the sequence information the lysozyme gene of the Klebsiella phage K11 was amplified and cloned using the polymerase chain reaction of the pfu DNA polymerase. The nucleotide sequence of phage K11 lysozyme gene was deter...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.03.026
更新日期:2005-07-01 00:00:00