Abstract:
:Staphylokinase (SAK) is reported to have a serine protease domain with no proteolytic activity unlike other plasminogen activators like tissue plasminogen activator (t-PA) and urokinase. A unique protease property of Staphylokinase was observed when SAK was expressed as a fusion protein in inducible Escherichia coli expression vectors. This finding was further investigated by cloning and expressing different SAK fusions, both native and N-terminal deletions, with fusion tags like glutathione S-transferase (GST) and signal sequence of SAK in bacterial system. While all the N-terminal SAK fusions were found to self-cleave in crude and purified preparations, the C-terminal SAK fusion was stable. The cleavage property of Staphylokinase fusion proteins, inhibited by reduced glutathione and PMSF, was independent of its thrombolytic activity and also independent on the type of host employed for its expression. The serine protease domain of the SAK gene possibly lies between 20th to 77th amino acid and serine 41 of this region appears critical for such a cleavage property.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Prasad B,Salunkhe SS,Padmanabhan Sdoi
10.1016/j.pep.2009.07.011subject
Has Abstractpub_date
2010-02-01 00:00:00pages
191-7issue
2eissn
1046-5928issn
1096-0279pii
S1046-5928(09)00179-Xjournal_volume
69pub_type
杂志文章abstract::The promyelocytic leukemia (PML) gene is involved in the 15/17 chromosomal translocation of acute promyelocytic leukemia (APL). It encodes a nuclear phosphoprotein containing an alpha-helical coiled-coil domain with four heptad repeats. The heptad repeats consist of four clusters of hydrophobic amino acids that mediat...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00004-4
更新日期:2003-05-01 00:00:00
abstract::Fusion proteins expressed in bacteria are often insoluble or inefficiently purified by standard procedures previously reported to work well for the non-fused proteins. We report here a simple but general procedure that can be used to quickly customize and optimize the purification of milligram quantities of most GST f...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00524-7
更新日期:2002-11-01 00:00:00
abstract::The high level expression and purification of rat monoamine oxidase B (rMAOB) in the methylotrophic yeast Pichia pastoris is reported. Nearly 100 mg of purified rMAOB is obtained from 130 g (wet weight) of cells (0.5 L of culture). The MALDI-TOF mass spectrum of the purified protein shows a single species with a molec...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.03.002
更新日期:2008-06-01 00:00:00
abstract::Hendra virus (family Paramyxoviridae) is a negative sense single-stranded RNA virus (NSRV) which has been found to cause disease in humans, horses, and experimentally in other animals, e.g. pigs and cats. Pteropid bats commonly known as flying foxes have been identified as the natural host reservoir. The Hendra virus ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.07.008
更新日期:2015-12-01 00:00:00
abstract::In microorganisms and plants, mevalonate kinase is involved in the biosynthesis of isoprenoid derivatives, one of the largest groups of natural products. We subcloned the gene of mevalonate kinase from Methanococcus jannaschii into a bacterial expression vector pLM1 with six continuous histidine codons attached to the...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00101-3
更新日期:2003-08-01 00:00:00
abstract::We have developed a family of cloning vectors that direct expression of fusion proteins that mimic aggregated immunoglobulin (IgG) (AIG) and immune complex function with respect to their interactions with FcgammaR and that allow for the inclusion and targeting of a second protein domain to cells expressing FcgammaR. T...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1406
更新日期:2001-04-01 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 human iron-binding protein lactoferrin (hLf) has been implicated in a number of important physiological pathways, including those regulating immune function and tumor growth. In an effort to develop an efficient system for production of recombinant hLf (rhLf) that is structurally and functionally equivalent to the...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.01.003
更新日期:2009-05-01 00:00:00
abstract::Mitochondrial 2,4-dienoyl-CoA reductase is a key enzyme for the beta-oxidation of unsaturated fatty acids. The cDNA of the full-length human mitochondrial 2,4-dienoyl-CoA reductase was previously cloned as pUC18::DECR. PCR methodologies were used to subclone the genes encoding various truncated human mitochondrial 2,4...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00191-8
更新日期:2003-10-01 00:00:00
abstract::Sarcotoxin IA is an antibacterial peptide that is secreted by a meat-fly Sarcophaga peregrina larva in response to a hypodermic injury or bacterial infection. This peptide is highly toxic against a broad spectrum of both Gram-positive and Gram-negative bacteria and lethal to microbes even at nanomolar concentrations. ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00697-6
更新日期:2003-04-01 00:00:00
abstract::Plantaricin JK (PlnJK) is a Class IIb LAB bacteriocin that includes two peptides; i.e., PlnJ and PlnK, which can synergistically halt many types of gram-positive bacteria, including food spoilage organisms. Purification of these peptides from natural lactic acid bacteria is difficult therefore, their application remai...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.02.013
更新日期:2019-07-01 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::Failure to express soluble proteins in bacteria is mainly attributed to the properties of the target protein itself, as well as the choice of the vector, the purification tag and the linker between the tag and protein, and codon usage. The expression of proteins with fusion tags to facilitate subsequent purification s...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.05.006
更新日期:2006-11-01 00:00:00
abstract::Periplasmic expression of recombinant proteins ensures the production of biologically active proteins in a correctly folded state with several key advantages. This research focused on the in-frame cloning of rhIL-15 in pET-20 (+) vector with pelB-leader sequence to direct the protein to the bacterial periplasm. The ta...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105746
更新日期:2021-01-01 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::Azotobacter vinelandii has recently been used for a variety of genetic experiments which take advantage of its facile transformation system and its high-frequency homologous recombination. One gene that has been cloned and sequenced is the fdxA gene that encodes a small Fe-S protein called A. vinelandii ferredoxin I (...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1014
更新日期:1994-02-01 00:00:00
abstract::The methylotrophic yeasts Pichia pastoris and Pichia angusta (Hansenula polymorpha) were used for the comparative heterologous production of two model mammalian proteins of pharmaceutical interest, the NK1-fragment (22 kDa) of human hepatocyte growth factor and the extracellular domain (28 kDa) of mouse tissue factor ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.03.010
更新日期:2009-08-01 00:00:00
abstract::Post-translational modification plays crucial roles in signal transduction in eukaryotic cells. To elucidate the biological function of a protein with a specific post-translational modification, it is necessary to isolate the modified protein. However, it is difficult to incorporate a modified amino acid into a specif...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.10.018
更新日期:2008-02-01 00:00:00
abstract::l-glutamate oxidase (GLOD), encoded by the gox gene, catalyses the transformation of l-glutamic acid into α-ketoglutaric acid (α-KG). In the present study, Pichia pastoris was used for heterologous production of GLOD following optimization of the gox coding sequence for expression in the yeast host. A series of constr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.09.014
更新日期:2017-01-01 00:00:00
abstract::Protein production can be improved if methods for soluble protein expression are developed. Interferon consensus (IFN-con) is used to treat hepatitis C. IFN-con has superior activity compared to other clinically used interferon α subtypes. However IFN-con is a challenging protein to produce in a soluble form using an ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.03.009
更新日期:2014-07-01 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::A new procedure for the large-scale purification of the recombinant thermostable chitinase (Chi40) cloned from Streptomyces thermoviolaceus in various expression vectors in Escherichia coli is described. Chi40 was overproduced in the cytosolic and secreted forms. The cytosolic form (Chi40c) was highly overproduced and...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1490
更新日期:2001-10-01 00:00:00
abstract::Baculovirus expression vectors are used routinely for foreign gene expression and are under intense development as improved biological pesticides. Conventional baculovirus expression vectors are recombinant viruses that can express a foreign gene in insect cells under the control of the polyhedrin promoter, which prov...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0092
更新日期:1996-09-01 00:00:00
abstract::Streptococcal protein G has been used extensively for the purification of antibodies using the interaction of the Fc region with protein G. Many antibodies also interact with protein G through a low-affinity binding site for the Fab region. The exploitation of this low-affinity interaction for the purification of Fab'...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(05)80037-3
更新日期:1992-10-01 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::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::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::For affinity-chromatography-based purification of proteins that are prone to abnormal termination of translation or that may not be modified at their N-termini, affinity tags are needed which can be fused to the C-terminus. In this publication we describe that maltose binding protein (MBP) fused to the C-terminus of t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0969
更新日期:1998-12-01 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::Lactobacillus β-galactosidases are mostly heterodimeric proteins, which are encoded by the two overlapping genes, lacL and lacM, and produced in recombinant prokaryotic systems for higher yield. This is the first report on the expression of a heterodimeric β-galactosidase from Lactobacillus crispatus B470 in Pichia pa...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.08.019
更新日期:2013-11-01 00:00:00