Purification, characterization, and crystallization of membrane bound Escherichia coli tyrosine kinase.

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

authors： Aksoy Y,Ogüs IH,Oauzer N

更新日期：2001-03-01 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

authors： Dolby V,Collén A,Lundqvist A,Cronet P

更新日期：2004-10-01 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

authors： Ye W,Liu J,Wang H,Wang J,Wang X

更新日期：2012-03-01 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

authors： Li Y,Han X,Xu W,Rao Z,Li X

更新日期：2019-06-01 00:00:00

abstract：:Cyclomaltodextrinases are multidomain and often dimeric proteins from the alpha-amylase family (glycoside hydrolase family 13) which frequently have been very difficult to express in active form in Escherichia coli. To express the soluble form of this type of proteins in larger quantities the expression has to be opti...

journal_title：Protein expression and purification

authors： Turner P,Holst O,Karlsson EN

更新日期：2005-01-01 00:00:00

abstract：:A stable mammalian cell line expressing highly active bovine interferon-gamma (BoIFN-γ) was generated using Flp recombinase-mediated integration. This recombinant 293 cell line (B1) efficiently secreted FLAG-tagged BoIFN-γ protein into the culture supernatant, as determined by ELISA and Western blot. The recombinant B...

journal_title：Protein expression and purification

authors： Xu Z,Shan F,Shan F,Meng C,Zhou X,Zhang X,Chen X,Jiao X

更新日期：2014-07-01 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

authors： Tang J,Hernández G,LeMaster DM

更新日期：2004-07-01 00:00:00

abstract：:Expression of recombinant proteins is an important step towards elucidating the functions of many genes discovered through genomic sequencing projects. It is also critical for validating gene targets and for developing effective therapies for many diseases. Here we describe a novel method to express recombinant protei...

journal_title：Protein expression and purification

authors： Wang A,Clapper J,Guderian JA,Foy TM,Fanger GR,Retter MW,Skeiky YA

更新日期：2003-07-01 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

authors： Cheung GL,Thomas TM,Rylatt DB

更新日期：2003-11-01 00:00:00

abstract：:S-locus protein kinase (SRK) is a receptor kinase that plays a critical role in self-recognition in the Brassicaceae self-incompatibility (SI) response. SRK is activated by binding of its ligand S-locus protein 11 (SP11) and subsequently induced phosphorylation of the intracellular kinase domain. However, a detailed a...

journal_title：Protein expression and purification

authors： Murase K,Hirano Y,Takayama S,Hakoshima T

更新日期：2017-03-01 00:00:00

abstract：:The SaeRS two-component system in Staphylococcus aureus controls the expression of a series of virulence factors, such as hemolysins, proteases, and coagulase. The response regulator, SaeR, belongs to the OmpR family with an N-terminal regulatory domain and a C-terminal DNA binding domain. To improve the production an...

journal_title：Protein expression and purification

authors： Fan R,Shi X,Guo B,Zhao J,Liu J,Quan C,Dong Y,Fan S

更新日期：2021-01-01 00:00:00

abstract：:Programmed cell death (apoptosis) is an essential mechanism in life that tightly regulates embryogenesis and removal of harmful cells. Besides an extrinsic pathway, an intrinsic (mitochondrial) apoptotic pathway exists where mitochondria are actively involved in cellular clearance in response to internal stress signal...

journal_title：Protein expression and purification

authors： Ådén J,Mushtaq AU,Dingeldein A,Wallgren M,Gröbner G

更新日期：2020-08-01 00:00:00

abstract：:BetaB2-crystallin, the major subunit of beta-crystallins, is difficult to purify either from lens homogenate or from betaH-or betaL-crystallins. It has been prepared by heterologous expression in Escherichia coli. Most often, the methods used for purifying a recombinant globular protein employ the combination of ion-e...

journal_title：Protein expression and purification

authors： Jobby MK,Sharma Y

更新日期：2003-03-01 00:00:00

abstract：:The aim of this study was to establish a method of purifying intact complement factor H (CFH) from human plasma. CFH was isolated from human plasma by polyethylene glycol (PEG) precipitation, following three sequential chromatographic columns, which consisted of l-lysine Sepharose column, Resource Q column and Sephacr...

journal_title：Protein expression and purification

authors： Wang FM,Yu F,Zhao MH

更新日期：2013-10-01 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

authors： Geisbrecht BV,Bouyain S,Pop M

更新日期：2006-03-01 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

authors： Dabrowski S,Olszewski M,Piatek R,Kur J

更新日期：2002-10-01 00:00:00

abstract：:A new method was developed to purify the streptococcal hyaluronate synthase in active form to electrophoretic homogeneity. The method is based on the extraction of protoplast membranes with digitonin and a phase separation into an aqueous and a detergent phase induced by addition of polyethylene glycol 6000 at 0 degre...

journal_title：Protein expression and purification

authors： Prehm S,Nickel V,Prehm P

更新日期：1996-06-01 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

authors： O'flaherty M,McMahon M,Mulcahy P

更新日期：1999-02-01 00:00:00

abstract：:Mannan outer chain N-glycan structures are yeast/fungal-specific typically found on secreted and cell wall glycoproteins. Mannan outer chains consist of an alpha-1,6 polymannose backbone attached to a Man(8-10)(GlcNAc)(2) core. The backbone contains branches of alpha-1,2 mannose residues, terminated with alpha-1,3 man...

journal_title：Protein expression and purification

authors： Rodionov D,Romero PA,Berghuis AM,Herscovics A

更新日期：2009-07-01 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

authors： Glynou K,Ioannou PC,Christopoulos TK

更新日期：2003-02-01 00:00:00

abstract：:Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor family that acts specifically on epithelial cells in a paracrine mode. We employed a mammalian expression system to synthesize recombinant human KGF and isolated two preparations, KGF-a and KGF-b, from medium conditioned by Chinese hamster ov...

journal_title：Protein expression and purification

authors： Hsu YR,Hsu EW,Katta V,Brankow D,Tseng J,Hu S,Morris CF,Kenney WC,Lu HS

更新日期：1998-03-01 00:00:00

abstract：:An alpha-amylase has been purified from the thermophilic fungus Scytalidium thermophilum. A ninefold purification was achieved in a single step using fluidized bed chromatography wherein alginate was used as the affinity matrix. There are at least two isoenzymes as shown by concanavalin A (Con A)-agarose column chroma...

journal_title：Protein expression and purification

authors： Roy I,Sastry MS,Johri BN,Gupta MN

更新日期：2000-11-01 00:00:00

abstract：:An efficient expression system for a low-molecular mass xylanase in Escherichia coli has been developed. A gene encoding the mature Bacillus circulans (Bc) xylanase was designed to imitate the frequency of degenerate codons used in E. coli. Appropriate degenerate codons were used to create multiple unique restriction ...

journal_title：Protein expression and purification

authors： Sung WL,Luk CK,Zahab DM,Wakarchuk W

更新日期：1993-06-01 00:00:00

abstract：:Splicing factor proline- and glutamine-rich (SFPQ) is an RNA-binding protein, playing significant roles in gene regulation and subnuclear body formation. Our recent serendipitous discovery showed that SFPQ binds zinc directly and forms an infinite polymer that is induced by zinc binding to the protein. The zinc-induce...

journal_title：Protein expression and purification

authors： Lim YW,Lee M

更新日期：2020-07-01 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

authors： Sharma M,Swain PK,Chopra AP,Chaudhary VK,Singh Y

更新日期：1996-02-01 00:00:00

abstract：:Hydrophobic interaction high-performance liquid chromatography (HIC-HPLC) was utilized for the separation of native human antithrombin (AT) and a partially denaturated form of AT, known as the latent form (L-AT). The AT used in this study is commercially available (Atenativ, Pharmacia & Upjohn, Sweden) and contains al...

journal_title：Protein expression and purification

authors： Karlsson G,Winge S

更新日期：2001-02-01 00:00:00

abstract：:The bivalent anti-human anti-T cell immunotoxin A-dmDT390-bisFv(G(4)S) was developed for treatment of T cell leukemia, autoimmune diseases, and tolerance induction for transplantation. The multi-domain structure of the bivalent immunotoxin hinders efficient production in Escherichia coli and most eukaryotes are sensit...

journal_title：Protein expression and purification

authors： Woo JH,Liu YY,Mathias A,Stavrou S,Wang Z,Thompson J,Neville DM Jr

更新日期：2002-07-01 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

authors： Ahmed N,Afroze B,Abbas R,Khan MA,Akram M,Tahir S,Bakht S,Munir A,Shahid AA

更新日期：2021-01-01 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

authors： Cipáková I,Hostinová E,Gasperík J,Velebný V

更新日期：2004-09-01 00:00:00

abstract：:Endostatin, a carboxy-terminal fragment of collagen XVIII, has been shown to act as an anti-angiogenic agent that specifically inhibits proliferation of endothelial cells and growth of various primary tumors. Here, we describe the expression by Chinese hamster ovary (CHO) cells of murine endostatin and of a tagged-fus...

journal_title：Protein expression and purification

authors： Chura-Chambi RM,Tornieri PH,Spencer PJ,Nascimento PA,Mathor MB,Morganti L

更新日期：2004-05-01 00:00:00