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 the protease activities of both trypsin and MMP1 to digest E. coli proteins and activate pro-MMP1. Identity of activated MMP1 was confirmed by Western blot using anti-human MMP1 antibodies, whereas the mass was determined to be 43 kD using matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF-MS). Collagen and gelatin degradation by purified MMP1 were confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) of degraded FITC-labeled type-1 collagen and gelatin zymogram. Broad-spectrum protease activity of purified MMP1 was also confirmed by lysis of native E. coli proteins. Inexpensive high throughput purification of recombinant human MMP1 in E. coli will enable easier MMP1 production for diverse applications.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Kumar L,Colomb W,Czerski J,Cox CR,Sarkar SKdoi
10.1016/j.pep.2018.04.001subject
Has Abstractpub_date
2018-08-01 00:00:00pages
59-67eissn
1046-5928issn
1096-0279pii
S1046-5928(18)30087-1journal_volume
148pub_type
杂志文章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：:We have developed a new expression vector, pcI(ts) ind(+), based upon the powerful rightward promoter of bacteriophage lambda, which is controlled by a temperature-sensitive and chemically-inducible version of the lambda repressor on the same plasmid. Locating the repressor gene on the plasmid makes this vector "porta...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.09.018
更新日期：2009-02-01 00:00:00
abstract：:This paper reports the overproduction and the details of a rapid method to purify active sigmaS monomers from a T7 RNA polymerase-based protein expression system. This 2-day procedure involves solubilizing inclusion bodies in sarkosyl detergent, removal of sarkosyl by dialysis, and a single gel filtration column chrom...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0069
更新日期：1996-08-01 00:00:00
abstract：:Structural studies of human peptide hormone somatostatin 14 (SS14) require high amounts of isotopically labelled SS14 to be produced. Here we report a method for effective production of isotopically labelled SS14. SS14 was expressed as a fusion protein with thioredoxin in Escherichia coli. Co-expression of a longer po...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.03.011
更新日期：2014-07-01 00:00:00
abstract：:The expression of recombinant proteins in bacterial hosts may alter the biophysical properties of the protein of interest as a result of differences in post-translational processing from that observed when produced in the native cell. For example, recombinant human interleukin-1beta (IL-1beta) is produced as three iso...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.01.010
更新日期：2005-05-01 00:00:00
abstract：:The membrane-anchored metalloproteinase ADAM17 (TNF-alpha converting enzyme; TACE; EC 3.4.24.86) continues to be an attractive drug target in inflammatory diseases and cancer. Cocrystallization of its catalytic domain with a lead compound was complicated by the tenacious retention of the prodomain that has been shown ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.10.021
更新日期：2007-04-01 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. The DNA sequence encoding the light chain (catalytic subunit) of human ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00159-1
更新日期：2003-09-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.09.016
更新日期：2019-02-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.04.005
更新日期：2008-08-01 00:00:00
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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：:Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is cr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.02.004
更新日期：2019-06-01 00:00:00
abstract：:Glycolate oxidase is a flavin-dependent enzyme in the photorespiratory pathway in plants. Here we report the heterologous expression of glycolate oxidase in Escherichia coli and an isolation procedure which results in 4 mg pure protein per gram cell paste in only 1.5 days. This corresponds to a more than 50-fold impro...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0103
更新日期：1996-11-01 00:00:00
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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：:Baculovirus mediated gene transduction of mammalian cells (BacMam) is an emerging technique for rapid recombinant protein expression in mammalian cells. We constructed two baculovirus transfer vectors that incorporate several mammalian transcriptional regulatory elements necessary for high-level protein expression in ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.08.004
更新日期：2008-12-01 00:00:00
abstract：:Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human bikunin (rh-bikunin) in the methylotrophic yeast Pichia pastoris and established the purification pr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.03.025
更新日期：2008-08-01 00:00:00
abstract：:We have investigated the suitability of Pichia pastoris as an expression system for the candidate therapeutic protein, Sonic hedgehog fused to an immunoglobulin Fc domain (Shh-Fc). Sonic hedgehog is a morphogen protein involved in the patterning of a wide range of tissues during animal embryogenesis. The presence of S...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00062-7
更新日期：2003-06-01 00:00:00
abstract：:Phenylalanine hydroxylase, important in phenylalanine metabolism in mammals, is regulated through short-term (activation) and long-term (induction) mechanisms. To help elucidate the structure-function relationships involved in the activation of this enzyme, we have isolated and characterized full-length cDNA clones to...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：
更新日期：1992-04-01 00:00:00
abstract：:General procedures for the rapid, large-scale purification of recombinant Lactobacillus casei thymidylate synthase and its mutants have been established. An effective method employs sequential phosphocellulose and hydroxylapatite chromatography. Crude cell extracts are directly applied to phosphocellulose, and the enz...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(05)80039-7
更新日期：1992-10-01 00:00:00
abstract：:Phthalate dioxygenase (PDO), a hexamer with one Rieske-type [2Fe-2S] and one Fe (II)-mononuclear center per monomer, and its reductase (PDR), which contains flavin mononucleotide and a plant-type ferredoxin [2Fe-2S] center, are expressed by Burkholderia cepacia at approximately 30mg of crude PDO and approximately 1mg ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.09.004
更新日期：2007-04-01 00:00:00
abstract：:Silent information regulator 2 (Sir2) proteins are a class of protein deacetylase enzymes that play key roles in transcriptional gene silencing, DNA repair, and aging. Here, we describe the high-level bacterial expression and purification of a human SirT2 construct that yields high resolution NMR spectra. By removing ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.12.006
更新日期：2006-07-01 00:00:00
abstract：:Aquaporins are integral membrane channel proteins found in all kingdoms of life. The Escherichia coli aquaporin Z (AqpZ) has been shown to solely conduct water at high permeability. Functional AqpZ is generally purified from the membrane fraction. However, the quantity of the purified protein is limited. In this study...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.08.014
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.105463
更新日期：2019-12-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.11.001
更新日期：2004-03-01 00:00:00
abstract：:Acyl carrier protein (ACP) was purified from Euglena gracilis variety bacillaris in yields of about 1 mg/100 g (wet wt) of cells. Antibodies against the purified protein were raised in hens and isolated from eggs. Antibodies raised against Euglena ACP inhibited the Euglena chloroplast nonaggregated fatty acid syntheta...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(91)90072-q
更新日期：1991-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1994.1050
更新日期：1994-08-01 00:00:00
abstract：:In this work, we present the production of an active 43 aa recombinant human beta-defensin-1 (rhBD-1(43)) in Escherichia coli AD202 cells using specific pLMM1-rhBD-1 expression system. Unique solubility properties of the C-terminal fragment of light meromyosin (LMM) allowed us to overcome foreseeable problems with iso...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.04.024
更新日期：2004-09-01 00:00:00
abstract：:A hexahistidine tag was fused to the N-terminus of apoaequorin. A suitable vector encoding the fusion protein was constructed and used for transformation of Escherichia coli JM109 cells. Apoaequorin was overexpressed under the control of tac promoter. It was found, however, that most of the protein existed in the form...
journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2003-02-01 00:00:00
abstract：:Enteroaggregative Escherichia coli (EAEC) heat-stable enterotoxin 1 (EAST1) is a 4.1k Da protein originally discovered in EAEC but known to be scattered in other diarrheagenic E. coli as well, possibly causing diarrhea in humans and animals. We report for the first time a method to express and purify EAST1 using the G...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.09.008
更新日期：2004-02-01 00:00:00
abstract：:The DEX gene encoding an extracellular dextranase from Lipomyces starkeyi was cloned into vector pPIC9k-His6 and was expressed in Pichia pastoris GS115 strain under the control of AOX1 promoter. After 107 h of the 5L-scaled fermentation, wet cells weight of the recombinant P. pastoris Mut(+) strain reached to 588.6g/L...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.10.021
更新日期：2008-03-01 00:00:00
abstract：:Peptide deformylases (PDFs) catalyze the removal of the formyl group from the N-terminal methionine residue in nascent polypeptide chains in prokaryotes. Its deformylation activity makes PDF an attractive candidate for the biocatalytic deprotection of formylated peptides that are used in chemoenzymatic peptide synthes...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2013.01.004
更新日期：2013-05-01 00:00:00
