当前位置： SCI文献检索 > PROTEIN EXPRESSION AND PURIFICATION期刊下所有文献 > Overexpression and functional characterisation of the human melanocortin 4 receptor in Sf9 cells.

Overexpression and functional characterisation of the human melanocortin 4 receptor in Sf9 cells.

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 MC4r corresponded to 0.25mg active receptor/litre culture volume. Addition of a viral signal peptide at the N-terminus of the His-tagged MC4r did not improve the expression level. Confocal laser microscopy studies revealed that both the N- and C-terminally tagged MC4r did not accumulate intracellularly and were mainly located in the plasma membrane. The recombinant receptors showed similar affinity for the agonist NDP-MSH (Kd = 11 nM) as to MC4r expressed in mammalian cells. Functional coupling of the highest expressed C-terminal tagged receptor to endogenous Galpha protein was demonstrated through GTPgammaS binding upon agonist stimulation of the receptor. Ki values for the ligands MTII, HS014, alpha-, beta-, and gamma-MSH are comparable to the values obtained for MC4r expressed in mammalian cells.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Dolby V,Collén A,Lundqvist A,Cronet P

doi

10.1016/j.pep.2004.06.018

keywords:

subject

Has Abstract

pub_date

2004-10-01 00:00:00

pages

455-61

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046592804002074

journal_volume

37

pub_type

杂志文章

相关文献

PROTEIN EXPRESSION AND PURIFICATION文献大全
  • Purification and biochemical characterization of recombinant rat liver phenylalanine hydroxylase produced in Escherichia coli.

    abstract：:Phenylalanine hydroxylase, important in phenylalanine metabolism in mammals, is regulated through short-term (activation) and long-term (induction) mechanisms. To help elucidate the structure-function relationships involved in the activation of this enzyme, we have isolated and characterized full-length cDNA clones to...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：

    authors： Citron BA,Davis MD,Kaufman S

    更新日期：1992-04-01 00:00:00

  • Formation of well-defined soluble aggregates upon fusion to MBP is a generic property of E6 proteins from various human papillomavirus species.

    abstract：:Protein aggregation is a main barrier hindering structural and functional studies of a number of interesting biological targets. The E6 oncoprotein of Human Papillomavirus strain 16 (E6(16)) is difficult to express under a native soluble form in bacteria. Produced as an unfused sequence, it forms inclusion bodies. Fus...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2006.07.029

    authors： Zanier K,Nominé Y,Charbonnier S,Ruhlmann C,Schultz P,Schweizer J,Travé G

    更新日期：2007-01-01 00:00:00

  • Generation and identification of variants with improved purification yield of Bowman-Birk protease inhibitors carrying protein binding loops.

    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

    authors： Collier KD,Vogtentanz G,Amin NS,Estabrook M,Estell DA,Fox B,Power SD,Rao R,Schmidt BF

    更新日期：2009-12-01 00:00:00

  • Expression and purification of a putative H-NS nucleoid-associated protein from the phytopathogen Xylella fastidiosa.

    abstract：:The H-NS protein is one of the major constituents of the nucleoid structure that has been implicated in the DNA packaging and in the global regulation of gene expression. The study of this transcriptional regulator is an effort to fight Xylella fastidiosa, a citrus pathogen responsible for a range of economically impo...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/S1046-5928(03)00193-1

    authors： Paula DP,Azzoni AR,Siqueira SF,Catani CF,Rosselli LK,de Souza AP

    更新日期：2003-11-01 00:00:00

  • Transmembrane-sequence-dependent overexpression and secretion of glycoproteins in Saccharomyces cerevisiae.

    abstract：:Protein expression using the secretory pathway in Saccharomyces cerevisiae can lead to high amounts of overexpressed and secreted proteins in culture supernatants in a short period of time. These post-translational modified expression products can be purified up to >90% in a single step. The overexpression and secreti...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1337

    authors： Schuster M,Wasserbauer E,Aversa G,Jungbauer A

    更新日期：2001-02-01 00:00:00

  • Design and preparation of non-tagged Yersinia pestis LcrV antigen in Escherichia coli and its immunogenicity in BALB/c mice.

    abstract：:The whole encoding sequence for Yersinia pestis LcrV antigen was cloned into pET-32a(+) and expressed in Escherichia coli BL21 (DE3). The LcrV was high level expressed in the E. coli cytoplasm in a completely soluble form. Recombinant LcrV could be purified from the supernatant of the bacteria lysate after chromatogra...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.10.011

    authors： li J,Li B,Li G,Ren J,Zhang J,Xu C,Yang X,Liu S,Fu L,Chen W

    更新日期：2008-02-01 00:00:00

  • Use of slow glucose feeding as supporting carbon source in lactose autoinduction medium improves the robustness of protein expression at different aeration conditions.

    abstract：:Recombinant protein expression from lac derived promoters by the autoinduction regime is based on diauxic growth of Escherichia coli on glucose and lactose. Glycerol is used as a supporting carbon source during the lactose-induced expression. While this glycerol-based formulation usually provides high cell densities, ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2013.07.016

    authors： Ukkonen K,Mayer S,Vasala A,Neubauer P

    更新日期：2013-10-01 00:00:00

  • Rapid generation of stable cell lines expressing corticotropin-releasing hormone receptor for drug discovery.

    abstract：:Human HEK293 cells that stably express the Epstein Barr nuclear antigen 1 (EBNA1) support the episomal replication of plasmids containing the Epstein Barr virus origin of replication (EBV oriP). A 293EBNA (293E) cell line expressing the human corticotropin-releasing hormone receptor subtype I (CRHR1) from an episomal ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0701

    authors： Horlick RA,Sperle K,Breth LA,Reid CC,Shen ES,Robbins AK,Cooke GM,Largent BL

    更新日期：1997-04-01 00:00:00

  • Targeting to the endoplasmic reticulum improves the folding of recombinant human telomerase reverse transcriptase.

    abstract：:Telomerase is a specialized reverse transcriptase that catalyzes the addition of telomeric repeats, TTAGGG in all vertebrates, to the ends of chromosomes. The lack of recombinant purified human telomerase reverse transcriptase (hTERT) has hampered biochemical and structural studies. The primary problem in generating a...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.05.016

    authors： Wu CK,Gousset K,Hughes SH

    更新日期：2007-11-01 00:00:00

  • Process development for production and purification of the Schistosoma mansoni Sm14 antigen.

    abstract：:The trematode Schistosoma mansoni Sm14 antigen was expressed in the yeast Pichia pastoris and secreted into the culture medium at yields of approximately 250 mg L-1. Sm14 belongs to a family of fatty-acid binding proteins and appears to play an important role in uptake, transport, and compartmentalization of lipids in...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2017.04.002

    authors： Damasceno L,Ritter G,Batt CA

    更新日期：2017-06-01 00:00:00

  • Amino-terminal charge affects the periplasmic accumulation of recombinant heregulin/EGF hybrids exported using the Escherichia coli alkaline phosphatase signal sequence.

    abstract：:An Escherichia coli expression system that exploits the bacterial alkaline phosphatase (PhoA) signal sequence to translocate recombinant human epidermal growth factor (hEGF) to the periplasm was used to evaluate how changes in the composition and sequence of amino acids near the PhoA-hEGF junction influence the peripl...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1997.0741

    authors： Campion SR,Elsasser E,Chung R

    更新日期：1997-08-01 00:00:00

  • Novel self-cleavage activity of Staphylokinase fusion proteins: An interesting finding and its possible applications.

    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

    authors： Prasad B,Salunkhe SS,Padmanabhan S

    更新日期：2010-02-01 00:00:00

  • Purification and biochemical characterization of the ErmSF macrolide-lincosamide-streptogramin B resistance factor protein expressed as a hexahistidine-tagged protein in Escherichia coli.

    abstract：:The erm proteins confer resistance to the MLS (macrolide-lincosamide-streptogramin B) antibiotics in various microorganisms, including pathogens, through dimethylation of a single adenine residue (A2085: Bacillus subtilis coordinate) of the 23S rRNA to reduce the affinity of antibiotics, thereby enabling the cells to ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2002.1621

    authors： Jin HJ,Yang YD

    更新日期：2002-06-01 00:00:00

  • Bacterial expression and antibiotic activities of recombinant variants of human β-defensins on pathogenic bacteria and M. tuberculosis.

    abstract：:Five variants of human β-defensins (HBDs) were expressed in Escherichia coli using two vector systems (pET28a(+) and pQE30) with inducible expression by IPTG. The last vector has not been previously reported as an expression system for HBDs. The recombinant peptides were different in their lengths and overall charge. ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2013.02.007

    authors： Corrales-Garcia L,Ortiz E,Castañeda-Delgado J,Rivas-Santiago B,Corzo G

    更新日期：2013-05-01 00:00:00

  • Production and characterization of biologically active human GM-CSF secreted by genetically modified plant cells.

    abstract：:Human granulocyte-macrophage colony-stimulating factor (GM-CSF), a hemopoietic growth factor, was produced and secreted from tobacco cell suspensions. The GM-CSF cDNA was carried by a binary vector under the control of the CaMV 35S promoter and the T7 terminator. In addition, a 5'-nontranslated region from the tobacco...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1232

    authors： James EA,Wang C,Wang Z,Reeves R,Shin JH,Magnuson NS,Lee JM

    更新日期：2000-06-01 00:00:00

  • Production and single-step purification of EGFP and a biotinylated version of the Human Rhinovirus 14 3C protease.

    abstract：:The fluorescent reporter enhanced Green Fluorescent Protein (EGFP) has been used for assaying a wide range of biological activities ranging from gene expression, or localization of target proteins through to intermolecular interactions. However, over-production of this protein in Escherichia coli has resulted in the p...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.05.003

    authors： Youell J,Fordham D,Firman K

    更新日期：2011-10-01 00:00:00

  • Improvement of extracellular secretion efficiency of Bacillus naganoensis pullulanase from recombinant Escherichia coli: Peptide fusion and cell wall modification.

    abstract：:Pullulanases are well-known starch-debranching enzymes that are widely used for hydrolysis of a-1,6-glycosidic linkages in starch, pullulan, amylopectin, and other oligosaccharides. Escherichia coli is a popular heterologous expression host for generating target enzymes. However, cells have to be disrupted to obtain t...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2018.09.011

    authors： Wang X,Chen Y,Nie Y,Xu Y

    更新日期：2019-03-01 00:00:00

  • Gag-derived proteins of HIV-1 isolates from Indian patients: cloning, expression, and purification of p24 of B- and C-subtypes.

    abstract：:A simple and efficient method for hyperexpression in Escherichia coli and purification of capsid protein, p24, of human immunodeficiency virus type 1 (HIV-1) of both B- and C-subtypes is described. DNA-encoding p24 of C-subtype was cloned from C-subtype gag sequence which was obtained by PCR amplification using DNA ex...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1266

    authors： Gupta S,Arora K,Gupta A,Chaudhary VK

    更新日期：2000-08-01 00:00:00

  • Purification of proteins with native terminal sequences using a Ni(II)-cleavable C-terminal hexahistidine affinity tag.

    abstract：:The role of the termini of protein sequences is often perturbed by remnant amino acids after the specific protease cleavage of the affinity tags and/or by the amino acids encoded by the plasmid at/around the restriction enzyme sites used to insert the genes. Here we describe a method for affinity purification of a met...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2019.03.009

    authors： Abd Elhameed HAH,Hajdu B,Balogh RK,Hermann E,Hunyadi-Gulyás É,Gyurcsik B

    更新日期：2019-07-01 00:00:00

  • Comparison of the expression of native and mutant bovine annexin IV in Escherichia coli using four different expression systems.

    abstract：:Bovine annexin IV, a Ca(2+)-dependent, membrane-binding protein, was expressed in E. coli using four different prokaryotic expression vector systems. An annexin IV cDNA was mutated in the 5' noncoding region to introduce an NcoI restriction site at the translation initiation site. The coding sequence was then excised ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1995.1016

    authors： Nelson MR,Creutz CE

    更新日期：1995-04-01 00:00:00

  • Functional Expression and Purification of CYP93C20，a Plant Membrane-Associated Cytochrome P450 from Medicago truncatula.

    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

    authors： Chang Z,Wang X,Wei R,Liu Z,Shan H,Fan G,Hu H

    更新日期：2010-12-04 00:00:00

  • A Protein Structure Initiative approach to expression, purification, and in situ delivery of human cytochrome b5 to membrane vesicles.

    abstract：:A specialized vector backbone from the Protein Structure Initiative was used to express full-length human cytochrome b5 as a C-terminal fusion to His8-maltose binding protein in Escherichia coli. The fusion protein could be completely cleaved by tobacco etch virus protease, and a yield of approximately 18 mg of purifi...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.11.018

    authors： Sobrado P,Goren MA,James D,Amundson CK,Fox BG

    更新日期：2008-04-01 00:00:00

  • Design and expression of polymeric immunoglobulin fusion proteins: a strategy for targeting low-affinity Fcgamma receptors.

    abstract：:We have developed a family of cloning vectors that direct expression of fusion proteins that mimic aggregated immunoglobulin (IgG) (AIG) and immune complex function with respect to their interactions with FcgammaR and that allow for the inclusion and targeting of a second protein domain to cells expressing FcgammaR. T...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1406

    authors： White DM,Jensen MA,Shi X,Qu Zx,Arnason BG

    更新日期：2001-04-01 00:00:00

  • Incorporating a TEV cleavage site reduces the solubility of nine recombinant mouse proteins.

    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

    authors： Kurz M,Cowieson NP,Robin G,Hume DA,Martin JL,Kobe B,Listwan P

    更新日期：2006-11-01 00:00:00

  • A modified clear-native polyacrylamide gel electrophoresis technique to investigate the oligomeric state of MBP-5-HT3A-intracellular domain chimeras.

    abstract：:The main principles of higher-order protein oligomerization are elucidated by many structural and biophysical studies. An astonishing number of proteins self-associate to form dimers or higher-order quaternary structures which further interact with other biomolecules to elicit complex cellular responses. In this study...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2018.08.010

    authors： Pandhare A,Stuebler AG,Pirayesh E,Jansen M

    更新日期：2019-01-01 00:00:00

  • Robust and facile purification of full-length, untagged human Nedd4 as a recombinant protein from Escherichia coli.

    abstract：:Nedd4 is an E3 ubiquitin ligase that has received increased attention due to its role in the maintenance of proteostasis and in cellular stress responses. Investigation of Nedd4 enzymology has revealed a complex enzymatic mechanism that involves intermolecular interactions with upstream E2 conjugating enzymes and with...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2020.105649

    authors： Hatstat AK,McCafferty DG

    更新日期：2020-09-01 00:00:00

  • High-level extracellular production of Rhizopus oryzae lipase in Pichia pastoris via a strategy combining optimization of gene-copy number with co-expression of ERAD-related proteins.

    abstract：:Rhizopus oryzae lipase (ROL) is an important industrial enzyme limited in application due to its low production in native strains. Here, we used a new combined strategy to overexpress ROL in Pichia pastoris. An efficient method based on bio-brick was developed to construct a series of vectors harboring different copy ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2018.02.005

    authors： Jiao L,Zhou Q,Su Z,Xu L,Yan Y

    更新日期：2018-07-01 00:00:00

  • Expression of soluble, biologically active recombinant human endostatin in Escherichia coli.

    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

    authors： Xu HM,Zhang GY,Ji XD,Cao L,Shu L,Hua ZC

    更新日期：2005-06-01 00:00:00

  • Cloning, purification, and refolding of human paraoxonase-3 expressed in Escherichia coli and its characterization.

    abstract：:Human paraoxonase (hPON3) is a high density lipoprotein-related glycoprotein with multi-enzymatic properties and antioxidant activity which is proposed to participate in the prevention of low density lipoprotein (LDL) oxidation. In this study, hPON3 gene was amplified from Human Fetal Liver Marathon-Ready cDNA and exp...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.07.021

    authors： Lu H,Zhu J,Zang Y,Ze Y,Qin J

    更新日期：2006-03-01 00:00:00

  • Efficient expression, purification and characterization of native human cystatin C in Escherichia coli periplasm.

    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

    authors： Zhou Y,Zhou Y,Li J,Chen J,Yao Y,Yu L,Peng D,Wang M,Su D,He Y,Gou L

    更新日期：2015-07-01 00:00:00