Abstract:
:The numerous proteins occluded within the molluscan shell play a key role in the control of the mineralization process. Although extensively studied, these proteins are still poorly known, mainly because they are difficult to fractionate. In the present paper, we present, for the first time, a simple combined strategy for separating successfully large amounts of molluscan shell proteins. Since shell proteins do not absorb at 280 nm, our approach is based on the "blind" separation of these proteins by a preparative denaturing electrophoresis. They are subsequently detected on dot-blot with polyclonal antibodies raised against the unfractionated soluble matrix. In the present case, this approach allows one to collect enough purified proteins to obtain amino-acid composition as well as N-terminal sequences, and to perform in vitro tests and glycosylation studies. Furthermore, this method permits one to raise polyclonal antibodies against the isolated proteins.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Marin F,Pereira L,Westbroek Pdoi
10.1006/prep.2001.1487keywords:
subject
Has Abstractpub_date
2001-10-01 00:00:00pages
175-9issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(01)91487-1journal_volume
23pub_type
杂志文章abstract::G-protein coupled receptors (GPCRs) are seven transmembrane helical proteins involved in cell signaling and response. They are targets for many existing therapeutic agents, and numerous drug discovery efforts. Production of large quantities of these receptors for drug screening and structural biology remains challengi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.10.017
更新日期:2007-04-01 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
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00075-5
更新日期:2003-07-01 00:00:00
abstract::Infections caused by Acinetobacter baumannii have emerged as a significant clinical problem due to the increase in infections caused by antibiotic resistant strains. A. baumannii OmpA is a highly conserved membrane protein that has multiple roles in interacting with the host during infection, and thus represents an at...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.11.019
更新日期:2011-05-01 00:00:00
abstract::Human interleukin-7 (IL-7) is a member of the interleukin family. Numerous studies have demonstrated IL-7's effect on B- and T-cell development as well as its potential in various clinical applications. Previously, a study reported that IL-7 could be purified from inclusion bodies using a prokaryotic system, however, ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.08.009
更新日期:2009-01-01 00:00:00
abstract::HSPC144 is a newly identified gene in human CD34(+) hematopoietic stem/progenitor cells. In this work, we have expressed and purified the 225-residue protein from Escherichia coli BL21 (DE3) and identified a stable fragment HSPC144-P (residues 44-225) by limited proteolysis method. The HSPC144-P fragment exhibits high...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.03.008
更新日期:2005-07-01 00:00:00
abstract::Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which is capable of selectively inducing apoptosis of cancer cells, is a potential targeted drug for cancer therapy. The TRAIL protein induces apoptosis only in trimeric form. However, the recombinant soluble TRAIL (sTRAIL) trimer has low stability and a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.08.004
更新日期:2015-11-01 00:00:00
abstract::Protein-based cellular therapeutics have been limited by getting molecules into cells and the fact that many proteins require accurate cellular localization for function. Cytoplasmic transduction peptide (CTP) is a newly designed transduction peptide that carries molecules across the cell membrane with a preference to...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.10.023
更新日期:2009-04-01 00:00:00
abstract::Advances in expressed protein ligation (EPL) methods that permit specific introduction of unique modifications into proteins have facilitated protein engineering, structure-function and protein interaction studies. An EPL-generated hybrid exchangeable apolipoprotein has been constructed from recombinant fragments of a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.04.002
更新日期:2007-08-01 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::ABCA4 is a member of the A subfamily of ATP-binding cassette transporters that consists of large integral membrane proteins implicated in inherited human diseases. ABCA4 assists in the clearance of N-retinylidene-phosphatidylethanolamine, a potentially toxic by-product of the visual cycle formed in photoreceptor cells...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.02.010
更新日期:2014-05-01 00:00:00
abstract::The entire extracellular domain of the human heat-stable enterotoxin (ST) receptor as well as a truncated N-terminal domain were cloned as glutathione S-transferase fusion proteins and expressed in Escherichia coli. The recombinant fusion proteins were purified from both the cytosol and the inclusion body fractions by...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0087
更新日期:1996-09-01 00:00:00
abstract::α-N-Acetylgalactosaminidase (α-GalNAc-ase; EC.3.2.1.49) is an exoglycosidase specific for the hydrolysis of terminal α-linked N-acetylgalactosamine in various sugar chains. The cDNA corresponding to the α-GalNAc-ase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Saccharomyces cerevisiae....
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.09.009
更新日期:2012-01-01 00:00:00
abstract::Stromal-cell-derived factor-1 (SDF-1alpha) is an 8-kDa chemokine that is constitutively expressed in bone-marrow-derived stromal cells and has been identified as a ligand for the CXCR4 receptor. We produced the chemokine recombinantly as methionine-SDF-1alpha in Escherichia coli without the leader peptide sequence. Th...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1402
更新日期:2001-04-01 00:00:00
abstract::Targeted anti-tumor toxins consist of a toxic functional moiety that is chemically linked or recombinantly fused to a cell-directing ligand. Ribosome-inactivating proteins (RIPs), especially type I RIPs such as saporin or dianthin, are commonly used as toxin components. Although expression of type I RIP-based fusion p...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.07.004
更新日期:2013-09-01 00:00:00
abstract::Replacing the chymotrypsin inhibitory loop of soybean Bowman-Birk inhibitor (sBBI) with a VEGF binding peptide (BBI-AV) significantly reduces the overall purification yield when BBI-AV is produced as a fusion protein in a Bacillussubtilis expression system. The low purification yield is primarily due to a higher fract...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.08.006
更新日期:2009-12-01 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::Plants possess very large numbers of biosynthetic cytochrome P450 enzymes. In spite of the importance of these enzymes for the synthesis of bioactive plant secondary metabolites, only two plant P450 structures has been obtained to date. Isoflavone synthase (IFS) is a membrane-associated cytochrome P450 enzyme catalyzi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.11.012
更新日期:2010-12-04 00:00:00
abstract::Hereditary enzymopenic methemoglobinemia is a rare disease that predominantly results from defects in either the erythrocytic (type I) or microsomal (type II) forms of the enzyme NADH:cytochrome b5 reductase (EC 1.6.2.2). All 25 currently identified type I and type II methemoglobinemia mutants have been expressed in E...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00046-9
更新日期:2003-07-01 00:00:00
abstract::A rabbit liver cDNA library in phage lambdagt10 was screened using the coding cDNA for human cytosolic serine hydroxymethyltransferase. A clone of 1754 bp was isolated and the nucleotide sequence showed an open reading frame of 1455 bp, which coded for rabbit cytosolic serine hydroxymethyltransferase and was flanked b...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0890
更新日期:1998-07-01 00:00:00
abstract::The HIV-1 Gag precursor protein, Pr55(Gag), is a multi-domain polyprotein that drives HIV-1 assembly. The morphological features of HIV-1 suggested Pr55(Gag) assumes a variety of different conformations during virion assembly and maturation, yet structural determination of HIV-1 Pr55(Gag) has not been possible due to ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.04.013
更新日期:2014-08-01 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
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abstract::An expression library was generated from a partial NcoI and HindIII digest of genomic DNA from the thermophilic bacterium, Bacillus stearothermophilus P1. The DNA fragments were cloned into the expression vector pQE-60 and transformed into Escherichia coli M15[EP4]. Sequence analysis of a lipase gene showed an open re...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1456
更新日期:2001-08-01 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::Escherichia coli tyrosine kinase (Etk) is a membrane bound kinase in gram-negative bacteria that regulates the export of capsular polysaccharides (CPS). The molecular mechanism behind CPS regulation remains unclear, despite access to a crystal structure of the cytoplasmic kinase domain of Etk. In this study, an effici...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.08.029
更新日期:2016-09-01 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::Antibodies specific to β-Glucocerebrosidase were selected from phage displayed naïve scFv libraries. Biopannings were performed against recombinant human protein β-Glucocerebrosidase immobilized on polystyrene surface, specific phages were eluted with 50% ethylene glycol in citrate buffer (pH 6.0). Several specific bi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105573
更新日期:2020-06-01 00:00:00
abstract::The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.08.008
更新日期:2015-11-01 00:00:00
abstract::Borneol is a bicyclic plant monoterpene. It can be degraded by soil microorganisms through the conversion of borneol dehydrogenase (BDH) and a known camphor degradation pathway. Recombinant BDH from Pseudomonas sp. TCU-HL1 was produced in the form of inclusion body. The refolded BDH1 tends to precipitate. Insoluble re...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105715
更新日期:2020-11-01 00:00:00
abstract::The immunosuppressive drug FK506 binds its targets FK506-binding protein (FKBP) family and modulates cellular processes. Recent studies demonstrated that FK506 shows anti-malaria effects. Newly identified FK506-binding protein 35 from Plasmodium falciparum (PfFKBP35) is assumed to be the molecular target of FK506 in t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.12.019
更新日期:2007-05-01 00:00:00
abstract::The present report documents an improved enzyme assay method for the mammalian L-alanine:4,5-dioxovalerate transaminase which is of significant utility in work with crude tissue homogenates, cell cultures, or purified enzyme preparations. We also describe a new and rapid purification procedure for this enzyme from rat...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1072
更新日期:1994-12-01 00:00:00