Abstract:
:Human GRO alpha, GRO beta, and GRO gamma are neutrophil chemoattractants structurally related to IL-8 and compete with IL-8 for binding to IL-8 receptors on neutrophils. These proteins are part of a large superfamily of chemotactic cytokines, the "chemokines," members of which share striking structural similarities. We have expressed GRO cDNA's in Escherichia coli as fusions to the MalE gene product, maltose-binding protein (MBP), in a way that allows separation of GRO and MBP moieties by factor Xa cleavage. GRO beta and GRO gamma expressed in bacteria were active in in vitro chemotaxis assays and were as effective as IL-8 in inducing chemotactic migration of neutrophils. Recombinant GRO beta was chemotactic rather than chemokinetic when tested by checkerboard analysis while GRO gamma showed evidence of chemokinetic as well as chemotactic activity. The activities of GRO beta and GRO gamma were not species-specific as both proteins were active on rat as well as human neutrophils and were inhibitable by antibodies raised against CINC, the rat GRO homolog. These data indicate that the MBP fusion protein expression system provides a rapid and simple method for obtaining large quantities of members of the chemokine protein family for biological uses.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Zagorski J,DeLarco JEdoi
10.1006/prep.1994.1050subject
Has Abstractpub_date
1994-08-01 00:00:00pages
337-45issue
4eissn
1046-5928issn
1096-0279pii
S1046-5928(84)71050-3journal_volume
5pub_type
杂志文章abstract::Rat granulocyte macrophage colony-stimulating factor (rGM-CSF) cDNA was amplified and cloned, and recombinant-rGM-CSF (R-rGM-CSF) was expressed and isolated from Escherichia coli. The synthesis of R-rGM-CSF was directed by a modified, inducible maltose binding protein (MBP) gene fusion expression vector, pMTR-23, and ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1077
更新日期:1995-10-01 00:00:00
abstract::Receptor for activated C-kinase (RACK1) binds to protein kinase C and functions as an anchor for several other cellular components. Most in vitro studies of RACK1 have been carried out with RACK1 fused to a soluble fusion protein partner, such as GST or MBP. Here, we show that fusion complexes may exist as large solub...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00135-9
更新日期:2003-09-01 00:00:00
abstract::The human melanocortin 4 receptor (MC4r) was successfully expressed in Sf9 cells using the baculovirus infection system. N- and C-terminally His-tagged receptors generated B(max) values of 14 and 23 pmol receptor/mg membrane protein, respectively. The highest expression level obtained with the C-terminally His-tagged ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.06.018
更新日期:2004-10-01 00:00:00
abstract::Expressed protein libraries are becoming a critical tool for new target discovery in the pharmaceutical industry. In order to get the most meaningful and comprehensive results from protein library screens, it is essential to have library proteins in their native conformation with proper post-translation modifications....
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.05.011
更新日期:2011-09-01 00:00:00
abstract::Autophagy is the process of degradation of intracellular proteins through the lysosome. Members of the tripartite motif (TRIM) proteins have shown to directly recognize autophagic cargo and also to act as a hub for the phagophore nucleation complex. The TRIM proteins are classically characterized by the presence of an...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.10.014
更新日期:2018-03-01 00:00:00
abstract::Vascular endothelial growth factor (VEGF) is one of the most significant mediators of angiogenesis, which interacts with a specific membrane receptor: VEGF receptor 2 (VEGFR2). Studies elsewhere have shown that, a VEGF-blocker can regulate several vital processes of tumor promotion. However, there is no literature evi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.04.010
更新日期:2013-08-01 00:00:00
abstract::A method was developed for the affinity purification of human complement properdin. The preparation is part of an integrated scheme in which over 20 human plasma proteins can be recovered in a highly purified form. The yield of properdin was 5.9 mg from 3 liters of plasma, amounting to a 28% recovery. The crucial step...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1026
更新日期:1994-04-01 00:00:00
abstract::A kappa-light chain from a Fab expression system was truncated by the insertion of a stop codon in the gene sequence to produce a variable light (VL) single domain antibody (dAb). Here, we describe the expression of dAb in the periplasm of Escherichia coli through fermentation in a defined media. Immunoglobulin bindin...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.07.013
更新日期:2007-02-01 00:00:00
abstract::CD23, a 45-kDa type II membrane glycoprotein present on B cells, monocytes, and other human immune cells, is a low-affinity receptor for IgE. The extracellular region of the membrane-bound human CD23 is processed into at least four soluble (s) CD23 forms, with apparent molecular masses of 37, 33, 29, and 25 kDa. High ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1448
更新日期:2001-07-01 00:00:00
abstract::Enteropeptidase (synonym: enterokinase, EC 3.4.21.9) is a heterodimeric serine protease of the intestinal brush border that activates trypsinogen by highly specific cleavage of the trypsinogen activation peptide following the sequence (Asp)(4)-Lys. It has also great biotechnological interest because of the unique subs...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.04.005
更新日期:2011-10-01 00:00:00
abstract::The kinetic locking-on strategy improves the selectivity of protein purification procedures based on immobilized cofactor derivatives through use of enzyme-specific substrate analogues in irrigants to promote biospecific adsorption. This paper describes the development and application of this strategy to the one-chrom...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0995
更新日期:1999-02-01 00:00:00
abstract::Human pancreatic ribonuclease (HP-RNase) has considerable promise as a therapeutic agent. Structure-function analyses of HP-RNase have been impeded by the difficulty of obtaining the enzyme from its host. Here, a gene encoding HP-RNase was designed, synthesized, and inserted into two expression vectors that then direc...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0036
更新日期:1996-05-01 00:00:00
abstract::A simple and efficient method for hyperexpression in Escherichia coli and purification of capsid protein, p24, of human immunodeficiency virus type 1 (HIV-1) of both B- and C-subtypes is described. DNA-encoding p24 of C-subtype was cloned from C-subtype gag sequence which was obtained by PCR amplification using DNA ex...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1266
更新日期:2000-08-01 00:00:00
abstract::Der p 2, a major allergen derived from the house dust mite Dermatophagoides pteronyssinus, is one of the most clinically relevant allergens worldwide. Recombinant Der p 2 (rDer p 2) is useful in clinical diagnosis and disease-specific immunotherapy. However, previous studies showed that Der p 2 can only be expressed i...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.01.012
更新日期:2016-05-01 00:00:00
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. ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1087
更新日期:1995-10-01 00:00:00
abstract::We examined expression of two plant genes encoding coclaurine N-methyltransferase (CMT) and norcoclaurine synthase (NCS) in Escherichia coli from the Salmonella entericaprpBCDE promoter (P(prpB)) and compared it to that from the strongest IPTG-inducible promoter, P(T7). In contrast to our previous study showing slight...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.06.008
更新日期:2008-10-01 00:00:00
abstract::The P(II) proteins comprise a family of widely distributed signal transduction proteins that integrate the signals of cellular nitrogen, carbon and energy status, and then regulate, by protein-protein interaction, the activity of a variety of target proteins including enzymes, transcriptional regulators and membrane t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.09.008
更新日期:2012-01-01 00:00:00
abstract::Escherichia coli oligoribonuclease (EcoORN), encoded by the orn gene, is a 3'-5' exonuclease that degrades short single-stranded oligoribonucleotides to rNMPs in the final step of RNA degradation. The orn gene is essential in E. coli, but not in higher organisms, and close homologues are present in other genomes from ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.10.005
更新日期:2008-02-01 00:00:00
abstract::Cystathionine beta-synthase (CBS) catalyzes the pyridoxal-50-phosphate-dependent condensation of L-serine and L-homocysteine to form L-cystathionine in the first step of the transsulfuration pathway. Although effective expression systems for recombinant human CBS (hCBS) have been developed, they require multiple chrom...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.10.012
更新日期:2009-04-01 00:00:00
abstract::Resistance to Inhibitors of Cholinesterase-8 (Ric-8) proteins are molecular chaperones that fold heterotrimeric G protein α subunits shortly after biosynthesis. Ric-8 proteins also act as test tube guanine nucleotide exchange factors (GEF) that promote Gα subunit GDP for GTP exchange. The GEF and chaperoning activitie...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.10.002
更新日期:2019-02-01 00:00:00
abstract::We have cloned, from total human liver RNA, the cDNA encoding apolipoprotein E3 (apoE3). Site-directed mutagenesis was used to obtain the cDNA encoding the apoE4 isoform, a major variant of this apolipoprotein in man. These two cDNAs were subcloned into the procaryotic expression vector pAHRS. A polyhistidine tag was ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0061
更新日期:1996-06-01 00:00:00
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::Formation of biominerals often involves specific proteins that modulate the process of matrix assembly, nucleation, and crystal growth. AP7 is an aragonite-associated protein of 7 kDa and is intrinsically disordered. The structural disorder of AP7 makes it very difficult to express in Escherchiacoli. In this work, we ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.05.002
更新日期:2014-08-01 00:00:00
abstract::Chloroplast transformation systems offer unique advantages in biotechnology, including high level of foreign gene expression, maternal inheritance, and polycistronic expression. We studied chloroplast expression of LTK63 (change Ser-->Lys at position 63 in the A subunit) which is the mutant of Escherichia coli heat-la...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.08.002
更新日期:2004-11-01 00:00:00
abstract::The Staphylococcus simulans gene encoding lysostaphin has been PCR amplified from pRG5 recombinant plasmid (ATCC 67076) and cloned into Escherichia coli expression pTYB12 vector (IMPACT-CN System, New England BioLabs) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag. The...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1454
更新日期:2001-08-01 00:00:00
abstract::A fibrinolytic enzyme was purified from the cultured mycelia of Armillaria mellea by ion-exchange chromatography followed by gel filtration, and was designated A. mellea metalloprotease (AMMP). The purification protocol resulted in a 627-fold purification of the enzyme, with a final yield of 6.05%. The apparent molecu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.05.004
更新日期:2005-09-01 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::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::Refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. We will review key parameters associated with (1) conformation of the protein solubilized from inc...
journal_title:Protein expression and purification
pub_type: 杂志文章,评审
doi:10.1016/s1046-5928(02)00641-1
更新日期:2003-03-01 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