Abstract:
:A specialized vector backbone from the Protein Structure Initiative was used to express full-length human cytochrome b5 as a C-terminal fusion to His8-maltose binding protein in Escherichia coli. The fusion protein could be completely cleaved by tobacco etch virus protease, and a yield of approximately 18 mg of purified full-length human cytochrome b5 per liter of culture medium was obtained (2.3mg per g of wet weight bacterial cells). In situ proteolysis of the fusion protein in the presence of chemically defined synthetic liposomes allowed facile spontaneous delivery of the functional peripheral membrane protein into a defined membrane environment without prior exposure to detergents or other lipids. The utility of this approach as a delivery method for production and incorporation of monotopic (peripheral) membrane proteins is discussed.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Sobrado P,Goren MA,James D,Amundson CK,Fox BGdoi
10.1016/j.pep.2007.11.018subject
Has Abstractpub_date
2008-04-01 00:00:00pages
229-41issue
2eissn
1046-5928issn
1096-0279pii
S1046-5928(07)00296-3journal_volume
58pub_type
杂志文章abstract::The basidiomycete Phanerochaete chrysosporium produces several isoforms of lignin peroxidase, which catalyzes the oxidative depolymerization of lignin To date, ion-exchange chromatography and preparative isoelectric focusing (IEF) have been commonly used for isolation of lignin peroxidase isoenzymes. In this work we h...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1044
更新日期:1995-06-01 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::An expression library was generated from a partial NcoI and HindIII digest of genomic DNA from the thermophilic bacterium, Bacillus stearothermophilus P1. The DNA fragments were cloned into the expression vector pQE-60 and transformed into Escherichia coli M15[EP4]. Sequence analysis of a lipase gene showed an open re...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1456
更新日期:2001-08-01 00:00:00
abstract::L-Threonine dehydrogenase was purified 10,000-fold to a specific activity approximately 300 mumol.min-1.mg-1 protein from porcine liver mitochondria. Purification to apparent homogeneity was achieved by sequential chromatography on DEAE Sepharose FF, Affi-Gel Blue, Sephacryl S-200, Matrex Gel Red A, and Matrex Gel Gre...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1061
更新日期:1994-10-01 00:00:00
abstract::The mRNA encoding the 51-kDa subunit of 6-phosphogluconate dehydrogenase (6PGDH) from sheep liver was reverse-transcribed and amplified. The resulting cDNA was reamplified in N-terminal and C-terminal segments and spliced to generate a full-length clone, and an internal cDNA fragment was also amplified. The full-lengt...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0896
更新日期:1998-07-01 00:00:00
abstract::Antimicrobial peptide LL-37 plays an important role in human body's first line of defense against infection. To better understand the mechanism of action, it is critical to elucidate the three-dimensional structure of LL-37 in complex with bacterial membranes. We present a bacterial expression system that allows the i...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.10.022
更新日期:2006-06-01 00:00:00
abstract::We report the molecular cloning, expression, and single-step homogeneous purification of a full-length asparaginyl tRNA synthetase (NRS) from Fasciola gigantica (FgNRS). Fasciola gigantica is a parasitic liver fluke of the class Trematoda. It causes fascioliasis that infects the liver of various mammals, including hum...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.10.009
更新日期:2018-03-01 00:00:00
abstract::Deoxynivalenol (DON), a mycotoxin produced by several Fusarium species, is a worldwide contaminant of food and feedstuffs. The DON-specific single-chain variable fragment (scFv) antibody was produced in recombinant Escherichia coli. The variable regions of the heavy chain (V(H)) and light chain (V(L)) cloned from the ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2003.12.008
更新日期:2004-05-01 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::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::The hepatitis E virus (HEV) capsid antigen has been proposed as a candidate subunit vaccine for the prevention of hepatitis E. The full-length HEV ORF2 protein product is predicted to contain 660 amino acids and to weigh 72,000 daltons. Expression of the HEV ORF2 capsid gene from recombinant baculoviruses in insect ce...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1997.0817
更新日期:1998-02-01 00:00:00
abstract::Dehydrodolichyl diphosphate synthase (DDPPs) catalyzes the sequential condensation of isopentenyl diphosphate with farnesyl diphosphate to synthesize long-chain dehydrodolichyl diphosphate, which serves as a precursor of glycosyl carrier in glycoprotein biosynthesis in eukaryotes. To perform kinetic and structural stu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1511
更新日期:2001-12-01 00:00:00
abstract::An unknown d-tagatose 3-epimerase (DTE) containing a IoIE domain was identified and cloned from Escherichia coli. This gene was subcloned into the prokaryotic expression vector pET-15b, and induced by IPTG in E. coli BL21 expression system. Through His-select gel column purification and fast-protein liquid chromatogra...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.06.015
更新日期:2015-10-01 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 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::Recombinant OSU VP4 protein, and outer capsid antigen of porcine rotavirus, was purified to a high level from the spent broth of baculovirus-infected Spodoptera frugiperda insect cells. Initial clarification of the broth with a 0-60% ammonium sulfate cut retained 93% of the total VP4. Q-sepharose ion exchange chromato...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1993.1030
更新日期:1993-06-01 00:00:00
abstract::The Gradiflow, a preparative electrophoresis instrument designed to separate molecules on the basis of their size and charge, was used to purify antibody Fab and F(ab')2 fragments. The method described is charge based, utilizing the difference in the pI between the antibody Fab/F(ab')2 fragments and antibody Fc fragme...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/S1046-5928(03)00219-5
更新日期:2003-11-01 00:00:00
abstract::This is the first report of an insect esterase efficiently expressed in the methylotrophic yeast Pichia pastoris (so far insect esterases have been produced only in the baculovirus system). Having isolated a Tribolium castaneum carboxylesterase cDNA (TCE), we were initially unable to express it in Escherichia coli or ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.04.011
更新日期:2005-08-01 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::Process development and optimization studies were performed in order to improve the purification process of (rhIFN-gamma). The objective was to generate material with higher purity and quantity. An in-process control screening was developed to obtain the optimal condition for column chromatographic purification by mea...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.07.002
更新日期:2007-02-01 00:00:00
abstract::Molecular-level investigation of proteins is increasingly important to researchers trying to understand the mechanisms of signal transmission. Heterotrimeric G proteins control the activation of many critical signal transmission cascades and are also implicated in numerous diseases. As part of a longer-term investigat...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2012.06.003
更新日期:2012-08-01 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::Telomerase is a specialized reverse transcriptase that catalyzes the addition of telomeric repeats, TTAGGG in all vertebrates, to the ends of chromosomes. The lack of recombinant purified human telomerase reverse transcriptase (hTERT) has hampered biochemical and structural studies. The primary problem in generating a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.05.016
更新日期:2007-11-01 00:00:00
abstract::Receptor tyrosine kinase like orphan receptor 2 (ROR2) is a co-receptor for some Wnt proteins including Wnt5a that activate the noncanonical Wnt/planar cell polarity (PCP) signaling pathway. Upregulation of ROR2 is associated with several cancer forms. The extracellular region of ROR2, which contains an immunoglobulin...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.02.015
更新日期:2019-06-01 00:00:00
abstract::Human interleukin-7 (IL-7) is a member of the interleukin family. Numerous studies have demonstrated IL-7's effect on B- and T-cell development as well as its potential in various clinical applications. Previously, a study reported that IL-7 could be purified from inclusion bodies using a prokaryotic system, however, ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.08.009
更新日期:2009-01-01 00:00:00
abstract::Human β-glucuronidase (GUS; EC 3.2.1.31) is a lysosomal enzyme that catalyzes the hydrolysis of β-d-glucuronic acid residues from the non-reducing termini of glycosaminoglycans. Impairment in GUS function leads to the metabolic disorder mucopolysaccharidosis type VII, also known as Sly syndrome. We produced GUS from a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.07.011
更新日期:2017-12-01 00:00:00
abstract::Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 is 60 amino acids long and lacks one of the three conserved disulfide bonds typically found in Kunitz domain modules. It binds specifically to voltage activated potassium channe...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.01.019
更新日期:2006-06-01 00:00:00
abstract::A simple scheme for the rapid and efficient isolation of rat pro-atrial natriuretic factor (pro-ANF) has been developed. An isolated rat adrenal cell bioassay for ANF was established to optimize heart tissue extraction and chromatography conditions. This assay is based on the ability of ANF to inhibit angiotensin II-s...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/1046-5928(90)90041-v
更新日期:1990-09-01 00:00:00
abstract::Soluble oligomers and fibrillar deposits of amyloid beta (Abeta) are key agents of Alzheimer's disease pathogenesis. However, the mechanism of amyloid aggregation and its interaction with live cells still remain unclear requiring the preparation of large amounts of pure and different Abeta peptides. Here we describe a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.02.009
更新日期:2009-07-01 00:00:00
abstract::The guanine nucleotide exchange factor EF-1beta gene from the thermoacidophilic archaeon Sulfolobus solfataricus (SsEF-1beta) was amplified by PCR and cloned into the pT7-7 expression vector. One of four selected clones harbored the T160C nucleotide substitution leading to the Y54H amino acid change in a hydrophobic r...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1997.0806
更新日期:1998-02-01 00:00:00