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 apolipoprotein E (apoE) and apolipophorin III (apoLp-III). A recombinant fusion protein comprised of human apoE N-terminal residues 1-111, a modified Saccharomyces cerevisiae intein and a chitin binding domain was subjected to 2-mercaptoethanesulfonic acid (MESNA) induced cleavage to generate apoE(1-111)-MESNA. A second fusion protein was comprised of a bacterial pelB leader peptide fused to a variant form of Galleria mellonella apoLp-III residues 1-91. The N-terminal pelB leader sequence directed the newly synthesized fusion protein to the Escherichia coli perisplamic space where endogenous leader peptidase cleavage generated the desired N-terminal cysteine-containing protein fragment. The resulting apoLp-III fragment, which contained no sequence tags or tails, escaped the bacteria and accumulated in the culture medium. When cultured in M9 minimal medium, Asp1Cys apoLp-III(1-91) was produced in high yield and was the sole major protein in the culture supernatant. Ligation reactions with apoE(1-111)-MESNA yielded an engineered hybrid apolipoprotein. The results document the utility of the pelB fusion protein system for generating active N-terminal cysteine containing proteins for EPL applications.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Hauser PS,Ryan ROdoi
10.1016/j.pep.2007.04.002subject
Has Abstractpub_date
2007-08-01 00:00:00pages
227-33issue
2eissn
1046-5928issn
1096-0279pii
S1046-5928(07)00095-2journal_volume
54pub_type
杂志文章abstract：:Plants produce a large number of cellulases that are either secreted or anchored in the plasma membrane where they likely function in various aspects of cellulose synthesis, modification and degradation during plant growth and development. Very few of these enzymes have been characterized in any detail, however. Here ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.08.009
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abstract：:We describe a single method to purify milligram amounts of authentic wild-type and mutant HIV-1 and HIV-2 Tat proteins overexpressed in Escherichia coli. This method takes advantage of the highly basic, positively charged RNA binding domain present in both HIV-1 and HIV-2 Tat, which also facilitated purification of HI...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0096
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0869
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00004-4
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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
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abstract：:Leishmaniasis is considered by the World Health Organization to be the second most important disease caused by a protozoan parasite. Biochemical and molecular biology studies can help in the understanding of the biology of the Leishmania parasite. All protozoan parasites, including Leishmania, are unable to synthesize...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.05.010
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abstract：:Rotavirus VP8* subunit is the minor trypsin cleavage product of the spike protein VP4, which is the major determinant of the viral infectivity and neutralization. To study the structure-function relationship of this fragment and to obtain type-specific reagents, substantial amounts of this protein are needed. Thus, fu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.09.014
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abstract：:High affinity interaction between S-protein and S-peptide fragments of bovine pancreatic RNase A has been recently used for construction of molecular vehicles for targeted drug delivery. The vehicle is assembled as a complex of drug carrier conjugated S-protein with S-peptide-tagged targeting protein. To avoid random ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00546-6
更新日期：2002-12-01 00:00:00
abstract：:Post-translational modification of proteins is a dynamic way of generating new protein-protein interaction interfaces that are critical for signaling networks in diverse cellular functions. Purified recombinant proteins frequently lack these signature modifications. Using the tumor suppressor p53 as the model protein,...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.01.015
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abstract：:Granulins (GRNs) are potent growth factors that are upregulated in many aggressive cancers from a wide range of organs. GRNs form tight, disulphide bonded, beta hairpin stacks, making them difficult to express in recombinant form. We recently described Ov-GRN-1, a GRN family member secreted by the carcinogenic liver f...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2011.06.018
更新日期：2011-10-01 00:00:00
abstract：:The recently classified group III trypsins include members like Atlantic cod (Gadus morhua) trypsin Y as well as seven analogues from other cold-adapted fish species. The eight group III trypsins have been characterized from their cDNAs and deduced amino acid sequences but none of the enzymes have been isolated from t...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.06.008
更新日期：2007-02-01 00:00:00
abstract：:Alphavirus nonstructural protein nsP1 possesses distinct methyltransferase (MTase) and guanylyltransferase (GTase) activities involved in the capping of viral RNAs. In alphaviruses, the methylation of GTP occurs before RNA transguanylation and nsP1 forms a covalent complex with m(7)GMP unlike the host mRNA guanylyltra...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2011.05.022
更新日期：2011-10-01 00:00:00
abstract：:Fatty acid desaturases are enzymes that introduce double bonds into fatty acyl chains, among which stearoyl-acyl carrier protein desaturase (S-ACP-DES) was widely distributed in the plant kingdom. We cloned the cDNA coding for fab2/ssi2, an S-ACP-DES from Arabidopsis thaliana, into the vector pET30a and heterologously...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.08.011
更新日期：2010-02-01 00:00:00
abstract：:Escherichia coli glutamic acid decarboxylase is a pyridoxal phosphate-dependent enzyme that catalyzes the alpha-decarboxylation of glutamate to yield 4-amino-butyrate and CO2. The E. coli chromosome contains two genes encoding for this enzyme, gadA and gadB, which map at distinct loci. Their protein products differ in...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0121
更新日期：1996-12-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.07.003
更新日期：2009-12-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.03.004
更新日期：2019-07-01 00:00:00
abstract：:A genomic DNA of 1416 bp containing an open reading frame encoding a manganese superoxide dismutase (Mn-SOD) from Tatumella ptyseos ct was cloned. Sequence analysis of this new gene revealed that it translates 205 amino acid residues. The deduced amino acid sequence showed variable identities (41-91%) with sequences o...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.10.003
更新日期：2005-03-01 00:00:00
abstract：:Structural biology places a high demand on proteins both in terms of quality and quantity. Although many protein expression and purification systems have been developed, an efficient and simple system which can be easily adapted is desirable. Here, we report a new system which combines improved expression, solubility ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.09.008
更新日期：2009-02-01 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：:Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is cr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.02.004
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.05.007
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1342
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.10.002
更新日期：2019-02-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.08.004
更新日期：2008-12-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.05.019
更新日期：2004-09-01 00:00:00
abstract：:The cofactor heme (Fe-protoporphyrin IX) plays many important roles in biology. Identification of novel proteins for the transport, chaperoning and delivery of heme in cells is of widespread interest. Here, we describe the use of heme conjugated magnetic beads for the isolation of heme-binding proteins from complex pr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2010.10.001
更新日期：2011-03-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.10.012
更新日期：2009-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.06.014
更新日期：2008-11-01 00:00:00
abstract：:This study describes comparison between IPTG and lactose induction on expression of caprine growth hormone (cGH), enhancing cell densities of Escherichia coli cultures and refolding the recombinant cGH, produced as inclusion bodies, to biologically active state. 2-3 times higher cell densities were obtained in shake f...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.05.011
更新日期：2009-11-01 00:00:00
abstract：:Human cystatin C (HCC), encoded by cystatin 3 gene, is a 13.3kDa endogenous cysteine proteinase inhibitor and an important biomarker of renal function. However, expressing recombinant cystatin C is difficult because of low yield and inclusion bodies in Escherichia coli (E. coli). In this study, we cloned HCC gene into...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.03.006
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