Abstract:
:Ricin A chain (RTA) mutants which had been modified by the addition of three lysine residues, three lysines and an alanine, or six histidine residues at the carboxyl terminus were expressed in Escherichia coli. The recombinant proteins were purified to homogeneity by ion-exchange chromatography on CM-Sepharose CL-6B. The 28S ribosomal RNA N-glycosidase activities of the three RTA mutants were indistinguishable from each other and from the activity of wild-type recombinant RTA. The RTA mutants were not impaired, compared with wild-type RTA, in their ability to reassociate with ricin B chain to form ricin holotoxin. Holotoxins containing mutant RTAs were as readily dissociated into subunits under reducing conditions as native holotoxin, and the RTA mutants were indistinguishable from wild-type RTA in the extent of their interaction with biological membranes. Ricin holotoxins containing the RTA mutants were, however, less cytotoxic to Vero cells than ricin containing wild-type RTA. At equivalent concentrations, a time course assay showed that holotoxin containing the mutant RTAs took longer to kill target cells than that containing wild-type recombinant RTA, suggesting that the mutant forms of RTA are less efficiently processed or translocated across an intracellular membrane than is wild-type RTA.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Simpson JC,Roberts LM,Lord JMdoi
10.1006/prep.1995.1087subject
Has Abstractpub_date
1995-10-01 00:00:00pages
665-70issue
5eissn
1046-5928issn
1096-0279pii
S1046-5928(85)71087-Xjournal_volume
6pub_type
杂志文章abstract::Glycine N-acyltransferase (GLYAT) is a phase II metabolic detoxification enzyme for exogenous (xenobiotic) and endogenous carboxylic acids; consisting of fatty acids, benzoic acid, and salicylic acid. GLYAT catalyzes the formation of hippurate (N-benzoylglycine) from the corresponding glycine and benzoyl-CoA. Herein, ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.02.007
更新日期:2014-05-01 00:00:00
abstract::Alginate lyase digestion is an efficient way to degrade alginate into oligosaccharides, which are useful in various areas including chemistry, pharmacy and food industry. Here we determined the sequence of Vibrio sp. QY102 sourced alginate lyase, and set up its heterologous expression in E. coli. We improved its secre...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.09.002
更新日期:2019-01-01 00:00:00
abstract::We have used Ni(2+)-affinity chromatography as a rapid and efficient method to purify a sensory rhodopsin I (SR-I) derivative containing six consecutive histidine residues at its C-terminus (His-tagged SR-I). The protein was expressed in Halobacterium salinarium by integrating the corresponding gene at the chromosomal...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.0009
更新日期:1995-12-01 00:00:00
abstract::Flt3 ligand (FL) is a potent hematopoietic cytokine that affects the growth and differentiation of hematopoietic progenitor and stem cells both in vivo and in vitro. Pichia pastoris transformants secreting high-level rhFL were obtained using 'yeastern blotting' method and the expression level in liquid was about 30 mg...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.04.001
更新日期:2005-08-01 00:00:00
abstract::The coding region for arginyl-tRNA synthetase from jack bean (Canavalia ensiformis) has been sequenced and cloned into the bacterial expression vector pET32a. Transformation of BL21 cells and induction with IPTG results in the high level expression of the protein fused N-terminally with thioredoxin and bearing a His-t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.05.008
更新日期:2008-10-01 00:00:00
abstract::Bovine pancreatic procarboxypeptidase A has been overexpressed in a soluble and activatable form in Escherichia coli. When the protein was expressed under the control of bacteriophage T7 promoter in E. coli ADA494 (a thioredoxin reductase deficient bacteria), a thioredoxin fusion protein was produced at relatively hig...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00573-9
更新日期:2003-02-01 00:00:00
abstract::The hexokinase (ATP;D-hexose 6-phosphotransferase, EC 2.7.1.1) of Schistosoma mansoni has been expressed in Escherichia coli as a fusion protein including an N-terminal polyhistidine tag and enterokinase cleavage site. The enzyme was purified by metal chelate chromatography to > 95% homogeneity, based on analysis by S...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0113
更新日期:1996-11-01 00:00:00
abstract::Five variants of human β-defensins (HBDs) were expressed in Escherichia coli using two vector systems (pET28a(+) and pQE30) with inducible expression by IPTG. The last vector has not been previously reported as an expression system for HBDs. The recombinant peptides were different in their lengths and overall charge. ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.02.007
更新日期:2013-05-01 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::PagP, a beta-barrel membrane protein found in Gram-negative bacteria, expresses robustly in inclusion bodies when its signal sequence is removed. We have developed a new fusion protein expression system based on PagP and demonstrated its utility in the expression of the unstructured N-terminal region of human cardiac ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2012.07.007
更新日期:2012-09-01 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::The Pseudomonas sp. have been long recognized for their exogenous lipolytic activities yet the genus still contains a lot of unexplored strains. Due to the versatile metabolic machinery and their potential for adaptation to fluctuating environmental conditions Pseudomonas sp. are of great interest for biotechnological...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.08.007
更新日期:2019-01-01 00:00:00
abstract::Plants possess very large numbers of biosynthetic cytochrome P450 enzymes. In spite of the importance of these enzymes for the synthesis of bioactive plant secondary metabolites, only two plant P450 structures has been obtained to date. Isoflavone synthase (IFS) is a membrane-associated cytochrome P450 enzyme catalyzi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.11.012
更新日期:2010-12-04 00:00:00
abstract::Introduction and expression of foreign genes in bacteria often results accumulation of the foreign protein(s) in inclusion bodies (IBs). The subsequent processes of refolding are slow, difficult and often fail to yield significant amounts of folded protein. RHG1 encoded by rhg1 was a soybean (Glycine max L. Merr.) tra...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.12.017
更新日期:2007-06-01 00:00:00
abstract::Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent. The aim of this study is to produce large quantities of highly pure and bioactive recombinant human TRAIL. Here, TRAIL was expressed in soluble form by pH-stat fed-batch cultivation and purified using a rapid and simple tw...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.06.007
更新日期:2016-10-01 00:00:00
abstract::The main principles of higher-order protein oligomerization are elucidated by many structural and biophysical studies. An astonishing number of proteins self-associate to form dimers or higher-order quaternary structures which further interact with other biomolecules to elicit complex cellular responses. In this study...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.08.010
更新日期:2019-01-01 00:00:00
abstract::Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria, archaea, and eukarya. We report here the identification, expression, and purification of the SSB-like proteins of the thermophilic bacteria Thermus thermophilus and T. aquaticus. The nucleotide (...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00504-1
更新日期:2002-10-01 00:00:00
abstract::Transcription regulation in the cell occurs in the context of chromatin. It follows that a thorough investigation of the mechanism of transcription regulation must take into account the role of chromatin structure. Through classical and molecular genetic experiments in yeast, great strides have been made in understand...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0716
更新日期:1997-06-01 00:00:00
abstract::Human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cDNA placed under the transcriptional control of polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus. Preliminary results of recombinant human prolactin expression as extracellular as we...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1419
更新日期:2001-07-01 00:00:00
abstract::Soybean root nodules possess a developmentally regulated acid phosphatase (ACP) that exhibits the highest specificity for purine 5'-nucleoside monophosphates. The enzyme is a glycosylated dimer of 28- and 31-kDa subunits, which appear to be products of the same gene but differ in posttranslational modifications. In or...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0935
更新日期:1998-10-01 00:00:00
abstract:INTRODUCTION:Glycodelin is a glycoprotein with different oligosaccharides that are responsible for its diverse biological functions in contraception and immunosuppression. Therefore, it is necessary to have access to adequate amounts of glycodelin with retained carbohydrate structure for functional studies because the ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.09.020
更新日期:2017-02-01 00:00:00
abstract::A putative laccase gene (lacG) from Geobacillus sp. JS12 was cloned and expressed as a fusion protein with six histidine residues in Escherichia coli BL21 (DE3) cells, and the protein was primarily found in inclusion bodies. The resulting insoluble proteins were solubilized with 6 M guanidine HCl and refolded using an...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105646
更新日期:2020-09-01 00:00:00
abstract::Replacing the chymotrypsin inhibitory loop of soybean Bowman-Birk inhibitor (sBBI) with a VEGF binding peptide (BBI-AV) significantly reduces the overall purification yield when BBI-AV is produced as a fusion protein in a Bacillussubtilis expression system. The low purification yield is primarily due to a higher fract...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.08.006
更新日期:2009-12-01 00:00:00
abstract::Human small nuclear (sn) RNA genes are transcribed by either RNA polymerase II or III depending upon the arrangement of their core promoter elements. Regardless of polymerase specificity, these genes share a requirement for a general transcription factor called the snRNA activating protein complex or SNAP(C). This mul...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.02.015
更新日期:2006-08-01 00:00:00
abstract::To facilitate studies of multicomponent protein complexes, I have developed an Escherichia coli expression system which coexpresses up to four polypeptides from a single plasmid. The modular nature of the system enables efficient subcloning of a gene into each of the 4 cassettes in the polycistronic expression vector....
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1363
更新日期:2001-02-01 00:00:00
abstract::Glucose-6-phosphate dehydrogenase was purified from human placenta using DEAE-Sepharose fast flow, 2',5'-ADP Sepharose 4B column chromatography, and chromatofocusing on PBE 94 with PB 74. The enzyme was purified with 62% yield and had a specific activity of 87 units per milligram protein. The pH optimum was determined...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1370
更新日期:2001-03-01 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::Insoluble expression of intrinsically soluble proteins with native activity is potentially a promising alternative to soluble expression of folded protein or insoluble expression of unfolded protein requiring refolding. Here, we attempted to express highly soluble halophilic His-rich metal binding protein (HP) as inso...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.01.001
更新日期:2019-04-01 00:00:00
abstract::Hsp47 is regarded as a collagen-specific chaperone with several suggested roles in collagen biosynthesis under normal and disease conditions. We describe here a procedure for the expression and purification of Hsp47 in Escherichia coli using the IMPACT expression system (New England Biolabs) where the guest gene is fu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1470
更新日期:2001-10-01 00:00:00
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
更新日期:2012-03-01 00:00:00