Abstract：
:We present a new streptavidin-binding peptide for both the purification and the detection of recombinant proteins. The peptide possesses nanomolar-affinity for streptavidin and therefore was termed Nano-tag. The Nano-tag(15) is 15 amino acids long and binds to streptavidin with a dissociation constant of 4 nM and the Nano-tag(9) is a 9-mer peptide with a dissociation constant of 17 nM. We demonstrate the one-step purification of Nano-tagged proteins, namely bovine heart fatty acid-binding protein (FABP), bacterial chloramphenicol acetyltransferase (CAT), and green fluorescent protein (GFP), from an in vitro translation system as well as from an Escherichia coli lysate. No significant influence of the Nano-tag(15) and of the conditions during affinity chromatography on maturation or activity of the proteins was observed whereas the Nano-tag(9) revealed a slight decline in the amount and activity of the synthesized proteins. The main advantage of the Nano-tag is the mild and specific elution with washing buffer plus biotin or related compounds, which enables the elution of the bound fusion protein from the streptavidin column in the native state. Additionally, the Nano-tag allowed the detection of recombinant proteins on Western blots by a streptavidin-alkaline phosphatase conjugate.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Lamla T,Erdmann VAdoi
10.1016/j.pep.2003.08.014keywords:
subject
Has Abstractpub_date
2004-01-01 00:00:00pages
39-47issue
1eissn
1046-5928issn
1096-0279pii
S1046592803002602journal_volume
33pub_type
杂志文章abstract：:The recombinant truncated endolysin LysK consisting of two catalytic domains, N-terminal CHAP and amidase-2 (LysKCA) was overexpressed in E. coli in the form of inclusion bodies (IBs). These IBs were dissolved in 6 M solution of urea followed by the refolding process. The refolding efficacy of the dilution and matrix-...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105683
更新日期：2020-10-01 00:00:00
abstract：:The hepatitis E virus (HEV) capsid antigen has been proposed as a candidate subunit vaccine for the prevention of hepatitis E. The full-length HEV ORF2 protein product is predicted to contain 660 amino acids and to weigh 72,000 daltons. Expression of the HEV ORF2 capsid gene from recombinant baculoviruses in insect ce...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0817
更新日期：1998-02-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105628
更新日期：2020-08-01 00:00:00
abstract：:We have successfully expressed and purified active human glycogen synthase-1 (hGYS1). Successful production of the recombinant hGYS1 protein was achieved by co-expression of hGYS1 and rabbit glycogenin (rGYG1) using the MultiBac baculovirus expression system (BEVS). Functional measurements of activity ratios of hGYS1 ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2013.05.007
更新日期：2013-08-01 00:00:00
abstract：:PagP, a beta-barrel membrane protein found in Gram-negative bacteria, expresses robustly in inclusion bodies when its signal sequence is removed. We have developed a new fusion protein expression system based on PagP and demonstrated its utility in the expression of the unstructured N-terminal region of human cardiac ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.07.007
更新日期：2012-09-01 00:00:00
abstract：:Cyclohexanone monooxygenase (CMO) is a soluble flavoenzyme originally isolated from Acinetobacter spp. which carries out Baeyer-Villiger reactions with cyclic ketone substrates. In the present study we cloned the Acinetobacter CMO gene and modified it for facile purification from heterologous expression systems by inc...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1340
更新日期：2001-02-01 00:00:00
abstract：:The major active form of human thrombin, alpha-thrombin, was analyzed by hydrophobic interaction high-performance liquid chromatography (HIC-HPLC). The chromatographic system included a polymeric phenyl column and elution was performed by a gradient, 2-0M sodium chloride (5-20 min). Total analysis time was 30 min per ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00596-x
更新日期：2003-01-01 00:00:00
abstract：:Chan Su is a traditional medicine prepared from toxic secretions from the auricular and skin glands of Chinese toads. Previous studies show that active components in Chan Su can inhibit the proliferation of tumor cells. To study the effect of Chan Su peptides on angiogenesis, fresh Chan Su was collected and its compon...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.105445
更新日期：2019-11-01 00:00:00
abstract：:Hepatitis C virus encodes two enveloped glycoproteins, E1 and E2, which are involved in viral attachment and entry into target cells. We have obtained in insect cells infected by recombinant baculovirus a chimeric secreted recombinant protein, E1(341)E2(661,) containing the ectodomains of E1 and E2. The described proc...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2010.02.012
更新日期：2010-06-01 00:00:00
abstract：:Endostatin, a 20kDa C-terminal fragment of collagen XVIII, is a potent anti-angiogenic protein and inhibitor of tumor growth. Recombinant endostatin was prepared from Escherichia coli deposited as insoluble, inactive inclusion bodies. In the present study, we produced soluble and biologically active recombinant human ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.09.021
更新日期：2005-06-01 00:00:00
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...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.04.001
更新日期：2018-08-01 00:00:00
abstract：:A mammalian expression vector with features optimized for simple expression and purification of secreted proteins has been developed. This vector was constructed to facilitate X-ray crystallographic studies of cysteine-rich glycoproteins that are difficult to express by other means. Proteins expressed with this vector...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1331
更新日期：2000-12-01 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：:Antimicrobial peptide LL-37 plays an important role in human body's first line of defense against infection. To better understand the mechanism of action, it is critical to elucidate the three-dimensional structure of LL-37 in complex with bacterial membranes. We present a bacterial expression system that allows the i...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.10.022
更新日期：2006-06-01 00:00:00
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：:Expression of recombinant proteins in bacterial or eukaryotic systems often results in aggregation rendering them unavailable for biochemical or structural studies. Protein aggregation is a costly problem for biomedical research. It forces research laboratories and the biomedical industry to search for alternative, mo...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.10.007
更新日期：2013-02-01 00:00:00
abstract：:A TRAIL-CM4 fusion protein in soluble form with tumor selective apoptosis and antibacterial functions was expressed in the Escherichia coli expression system and isolated through dialysis refolding and histidine-tag Nickel-affinity purification. Fresh Jurkat cells were treated with the TRAIL-CM4 fusion protein. Trypan...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2016.02.015
更新日期：2016-06-01 00:00:00
abstract：:Fab fragments isolated from papain digests of monoclonal antibodies have a wide variety of uses in analytical and in both in vivo and in vitro diagnostic applications. A novel, non-affinity method which uses the Gradiflow to purify Fab fragments from the papain digest of a mouse IgG1 anti-c-myc monoclonal antibody is ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.07.005
更新日期：2003-12-01 00:00:00
abstract：:Membrane proteins, including solute transporters play crucial roles in cellular function and have been implicated in a variety of important diseases, and as such are considered important targets for drug development. Currently the drug discovery process is heavily reliant on the structural and functional information d...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.07.003
更新日期：2014-09-01 00:00:00
abstract：:Influenza A virus displays one of the highest infection rates of all human viruses and therefore represents a severe human health threat associated with an important economical challenge. Influenza matrix protein 2 (M2) is a membrane protein of the viral envelope that forms a proton selective ion channel. Here we repo...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2016.11.001
更新日期：2017-03-01 00:00:00
abstract：:The gene coding for phosphoglucomutase (PGM) from Oryctolagus cuniculus (rabbit) has been expressed in Escherichia coli under a T7 expression system with a His-tag. About half of the expressed PGM protein was present in inclusion bodies, but this protein was inactive when solubilized. The protein in the soluble cell f...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1288
更新日期：2000-10-01 00:00:00
abstract：:The full-length cDNA that encodes the hemolytic toxin Avt-I, with 226 amino acids, from the venomous sea anemone Actineria villosa has been cloned using the oligo-capping method. The cDNA contains 681bp open reading frame and its predicted amino acid sequences revealed that Avt-I was basic polypeptides without cystein...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.12.003
更新日期：2005-04-01 00:00:00
abstract：:The Ig-binding properties of protein L from Peptostreptococcus magnus and protein G from Streptococcus have been successfully combined through the construction of a novel hybrid protein, consisting of a single Ig-binding domain from each protein. The biophysical and biochemical properties of this construct have been c...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.11.007
更新日期：2008-03-01 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 TATA-binding protein (TBP) plays a central role in the assembly of most eukaryotic transcription initiation complexes. We have characterized 3 monoclonal antibodies (mAbs) that react in the far amino-terminal (N-terminal) domain of the human TBP molecule (residues 1-99). One of these mAbs (designated 1TBP22) is a ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.02.020
更新日期：2004-08-01 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：:To facilitate studies of multicomponent protein complexes, I have developed an Escherichia coli expression system which coexpresses up to four polypeptides from a single plasmid. The modular nature of the system enables efficient subcloning of a gene into each of the 4 cassettes in the polycistronic expression vector....
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1363
更新日期：2001-02-01 00:00:00
abstract：:A full-length cDNA sequence of plant type CRY (designated Hae-P-CRY) was cloned from the green alga Haematococcus pluvialis. The cDNA sequence was 3608 base pairs (bp) in length, which contained a 2988-bp open reading frame encoding 995 amino acids with molecular mass of 107.7 kDa and isoelectric point of 6.19. Multip...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105633
更新日期：2020-08-01 00:00:00
abstract：:Using a codon-optimized gene fragment, we report remarkable yields for extracellular domain of human NK cell receptor (NKp30ex) when produced on M9 minimal medium, even with low (2g/L) glucose concentration. The yields were identical using media containing (15)NH(4)Cl or (15)NH(4)Cl in combination with all-(13)C-d-glu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.09.016
更新日期：2012-12-01 00:00:00
abstract：:The AP-1 transcription factor is a dimeric protein complex formed primarily between Jun (c-Jun, JunB, JunD) and Fos (c-Fos, FosB, Fra-1, Fra-2) family members. These distinct AP-1 complexes are expressed in many cell types and modulate target gene expression implicated in cell proliferation, differentiation, and stres...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.01.016
更新日期：2008-05-01 00:00:00
