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 Green A. The subunit molecular mass was estimated as 37 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, while an apparent native molecular mass of 73 kDa was shown by gel filtration chromatography, suggesting a dimeric structure. The purified enzyme was subjected to proteolytic degradation and the resulting peptides were isolated exclusively by reverse-phase high-performance liquid chromatography. Approximately 70% of the total sequence was obtained and the N-terminal amino acid sequence of the intact polypeptide chain was thus tentatively extended to residue 40.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Kao YC,Davis Ldoi
10.1006/prep.1994.1061subject
Has Abstractpub_date
1994-10-01 00:00:00pages
423-31issue
5eissn
1046-5928issn
1096-0279pii
S1046-5928(84)71061-8journal_volume
5pub_type
杂志文章abstract：:Pen b 26 is one of the allergens produced by Penicillium brevicompactum which is one of the most prevalent in door airborne fungi and a major source of respiratory problems, including asthma. Pen b 26 wa scloned and expressed as an N-terminal as well as a C-terminal His6 tagged fusion protein in Escherichia coli. This...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.01.004
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abstract：:Androgen receptor (AR)-associated coregulator 70 (ARA70) is a cytoplasmic protein that has been characterized to have the ability to induce AR transcriptional activity in response to androgens and anti-androgens in prostate cancer cells. AR has been shown to have an important role in the progression of prostate cancer...
journal_title：Protein expression and purification
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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
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.02.004
更新日期：2015-05-01 00:00:00
abstract：:Transmembrane and coiled-coil domains 1 (TMCO1) has a highly conserved amino acid sequence among species, indicating a critical role of TMCO1 in cell physiology. The deficiency of TMCO1 in humans is associated with cerebrofaciothoracic dysplasia (CFTD), glaucoma, osteogenesis and the occurrence of cancer. TMCO1 was re...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105803
更新日期：2021-03-01 00:00:00
abstract：:(S)-Hydroxynitrile lyase (Hnl) from the tropical rubber tree Hevea brasiliensis catalyzes the formation of (S)-cyanohydrins from hydrocyanic acid and aldehydes or ketones. This enzyme accepts aliphatic, aromatic, and heterocyclic carbonyl compounds as substrates and is therefore considered a potent biocatalyst for the...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0765
更新日期：1997-10-01 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
pub_type： 杂志文章
doi：10.1006/prep.1996.0051
更新日期：1996-06-01 00:00:00
abstract：:The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector. Methods were developed to overexpress the TyrR protein of H. influenzae in Escherichia coli and to purify the protein on a large scale. Both in...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0757
更新日期：1997-07-01 00:00:00
abstract：:Rab GTPases are post-translationally geranylgeranylated on their C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase) and this modification is essential for their biological activity. Rab Escort Protein (REP) is a molecular chaperone that assists in the prenylation reaction carried out by RabGGTase. Muta...
journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2002-10-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.08.025
更新日期：2004-01-01 00:00:00
abstract：:Heparinase I (HepA) was originally isolated from Flavobacterium heparinum (F. heparinum) and specifically cleaves heparin/heparan sulfate in a site-dependent manner, showing great promise for producing low molecular weight heparin (LMWH). However, expressing recombinant HepA is extremely difficult in Escherichia coli ...
journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2012-06-01 00:00:00
abstract：:Wild-type and deglycosylated forms of human prostate-specific antigen were expressed in Chinese hamster ovary (CHO) cells as zymogens. ProPSA was collected from conditioned medium and purified using a single cation-exchange chromatographic step for the deglycosylated form and cation-exchange followed by gel filtration...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1342
更新日期：2000-12-01 00:00:00
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 e...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.07.011
更新日期：2010-02-01 00:00:00
abstract：:The high specific lysyl endopeptidase (Lys-C; EC 3.4.21.50) is often used for the initial fragmentation of polypeptide chains during protein sequence analysis. However, due to its specificity it could be a useful tool for the production of tailor-made protein hydrolysates with for example bioactive or techno functiona...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.09.024
更新日期：2016-02-01 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：:The sequence and structure of the target protein exert a marked effect on its soluble expression in Escherichia coli. The effects of the mutation of an amylase isolated from Bacillus licheniformis (BLA) on its soluble expression in E. coli were investigated. A random mutation library of BLA was constructed to screen f...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.12.010
更新日期：2016-04-01 00:00:00
abstract：:Three human cytochrome P450 1A1 (CYP1A1) allelic variants, namely wild-type (CYP1A1.1), CYP1A1.2 (I462V), and CYP1A1.4 (T461N), were expressed as C-terminal His-tagged fusions including a thrombin cleavage site in Spodoptera frugiperda insect cells by baculovirus infection. The variants were expressed with 30-90 nmol ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00672-1
更新日期：2003-04-01 00:00:00
abstract：:The iron-sulfur protein subunit, known as the Rieske protein, is one of the central components of the cytochrome b(6)f complex residing in chloroplast and cyanobacterial thylakoid membranes. We have constructed plasmids for overexpression in Escherichia coli of full-length and truncated Rieske (PetC) proteins from the...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00016-0
更新日期：2003-05-01 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：: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：:The introduction of an affinity tag offers an attractive approach to isolation of membrane proteins. The type of affinity tag and its positioning in the protein is determined by the desired subsequent experimental uses of the isolated protein. To minimize the risk of interference, membrane proteins may preferentially ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.06.005
更新日期：2009-11-01 00:00:00
abstract：:Protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. It is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. The determi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.06.014
更新日期：2004-10-01 00:00:00
abstract：:The assembly pathway of the insect cell Spodoptera frugiperda (Sf-9) was engineered to include expression of the murine chaperone immunoglobulin heavy chain binding protein (BiP) using the baculovirus vector. The impact of BiP coexpression on the production and secretion of functional and soluble recombinant immunoglo...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1994.1082
更新日期：1994-12-01 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：:To study the subunit structure and the active site of human immunodeficiency virus reverse transcriptase (RT), the enzyme was expressed in E. coli and purified to homogeneity in large quantities. The recombinant enzyme consists of two major polypeptides of 66,000 and 53,000 Da in equimolar amounts and a minor species ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(92)90005-h
更新日期：1992-08-01 00:00:00
abstract：:The Plasmodium falciparum cysteine protease falcipain-2 is a potential new target for antimalarial chemotherapy. In order to obtain large quantities of active falcipain-2 for biochemical and structural analysis, a systematic assessment of optimal parameters for the expression and refolding of the protease was carried ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1416
更新日期：2001-06-01 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
pub_type： 杂志文章
doi：10.1006/prep.2000.1349
更新日期：2001-02-01 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：:Large amounts of soluble fibronectin were easily purified from cryoprecipitated or fresh citrated human blood plasma by a three-step combination of gelatin and heparin-cellufine affinity chromatography. The elution conditions were optimized to obtain a homogeneous fraction on SDS-PAGE and Western blot under reducing c...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1103
更新日期：1999-10-01 00:00:00
abstract：:The leucine aminopeptidase from Aeromonas proteolytica (also known as Vibrio proteolyticus) (AAP) is a metalloenzyme with broad substrate specificity. The open reading frame (ORF) for AAP encodes a 54 kDa enzyme, however, the extracellular enzyme has a molecular weight of 43 kDa. This form of AAP is further processed ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.05.004
更新日期：2004-10-01 00:00:00
