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Protein Engineering (3):
Recent article
Protein Engineering
Journal of Cell Science (124)Pt 16 2011
Abstract
We provide novel insights into the roles of the CB2 splice variants, CB2(SH3+) and CB2(SH3-), and their substrate, Cdc42, in regulating gephyrin clustering at GABAergic synapses. Transfection of Myc-tagged CB2(SH3+) and CB2(SH3-) into cultured neurons revealed strong, but distinct, effects promoting...
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PMID: 21807943
PDF is available here.
PDF is available here.
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Abstract
We previously reported (Koshi, Y.; Nakata, E.; Miyagawa, M.; Tsukiji, S.; Ogawa, T.; Hamachi, I. J. Am. Chem. Soc.2008, 130, 245.), here we have developed a new organocatalyst-based approach that allows specific chemical acylation of a receptor protein on the surface of live cells. The catalysts con...
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PMID: 21761940
PDF is available here.
PDF is available here.
Abstract
We investigated the function and calcium regulation of the COOH-terminal end segment of TnI. A TnI model molecule was labeled with Alexa fluor 532 at a Cys engineered at the COOH-terminal end and used to reconstitute tertiary troponin complex. A Ca(2+) -regulated conformational change in the COOH te...
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PMID: 21777381
PDF is available here.
PDF is available here.
Glycoprotein labeling using engineered variants of galactose oxidase obtained by directed evolution.
Abstract
A directed evolution approach has been used for the generation of variants of galactose oxidase (GOase) that can selectively oxidize glycans on glycoproteins. The aldehyde function introduced on the glycans d-mannose (Man) and d-N-acetyl glucosamine (GlcNAc) by the enzyme variants could then be used...
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PMID: 21526835
PDF is available here.
PDF is available here.
Abstract
We reported the development of a clickable SAM analogue cofactor, 4-propargyloxy-but-2-enyl SAM, and its implementation to label substrates of human protein arginine methyltransferase 1 (PRMT1). In the system, the SAM analogue cofactor, coupled with matched PRMT1 mutants rather than native PRMT1, wa...
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PMID: 21539310
PDF is available here.
PDF is available here.
Xenobiotica (41)5 2011
Abstract
Many biosensors depend on molecular recognition reagents to achieve highly sensitive and specific detection levels of an analyte of interest. Although new and improved detection platforms continue to be developed, improvements in the affinity and specificity of the molecular recognition reagents oft...
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PMID: 19159106
PDF is available here.
PDF is available here.
Xenobiotica (41)5 2011
Abstract
Microbodies are novel pharmacophoric entities which are derived from naturally occurring cystine-knot microproteins. They provide extremely stable scaffolds that can be engineered to high-affinity binding proteins. A peptide-grafting approach yielded specific ligands for human thrombopoietin recepto...
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PMID: 19377985
PDF is available here.
PDF is available here.
Abstract
The incidence of hepatocellular carcinoma (HCC) in China is closely related to the population infected with hepatitis B virus (HBV). HCC cells with HBV secrete soluble HBsAg into blood but do not express it on the cell membrane. This study aimed to construct and investigate a new glycosyl-phosphatidy...
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PMID: 21459723
PDF is available here.
PDF is available here.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] (27)2 2011
Abstract
To understand the effectiveness of prokaryotic expression of fusion protein (F) of human metapneumovirus (hMPV) and its application as antigen, F proteins from different genotypes of hMPV were expressed in prokaryotic expression system and purified by Ni-NTA affinity chromatography column. According...
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PMID: 21528539
PDF is available here.
PDF is available here.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] (27)2 2011
Abstract
Abstract
A novel kind of fluorescent protein relying on the intramolecular interplay between two different fluorophores, one of chemical origin and one of biological origin, was developed. The fluorescent non-natural amino acid l-(7-hydroxycoumarin-4-yl)ethylglycine was site-specifically incorporated into the...
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PMID: 21341705
PDF is available here.
PDF is available here.
Abstract
We describe a method to map the active site configuration of engineered P450 variants in high throughput using a set of semisynthetic chromogenic probes. Through analysis of the resulting 'fingerprints', reliable predictions can be made regarding the reactivity of these enzymes toward complex substr...
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PMID: 21341707
PDF is available here.
PDF is available here.
Abstract
Alginate lyase enzymes represent prospective biotherapeutic agents for treating bacterial infections, particularly in the cystic fibrosis airway. To effectively deimmunize one therapeutic candidate while maintaining high level catalytic proficiency, a combined genetic engineering-PEGylation strategy...
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PMID: 21340021
PDF is available here.
PDF is available here.
Abstract
We show that the CatD ortholog of the budding yeast Saccharomyces cerevisiae (Pep4p) harbors a dual cytoprotective function, composed of an anti-apoptotic part, conferred by its proteolytic capacity, and an anti-necrotic part, which resides in the protein's proteolytically inactive propeptide. Thus,...
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PMID: 21593793
PDF is available here.
PDF is available here.
Abstract
Recombinant monoclonal antibodies (mAB) are a new class of medicinal preparations with the potential for wide application to the treatment of viral infections. Marked progress in the development of therapeutic mAB against respiratory syncytial influenza A viruses, West Nile virus, and some others has...
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PMID: 21786598
PDF is available here.
PDF is available here.
Abstract
The 6th European Antibody Congress (EAC), organized by Terrapinn Ltd., was held in Geneva, Switzerland, which was also the location of the 4th and 5th EAC. As was the case in 2008 and 2009, the EAC was again the largest antibody congress held in Europe, drawing nearly 250 delegates i...
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PMID: 21441785
PDF is available here.
PDF is available here.
Abstract
The 21st Annual Antibody Engineering and 8th Annual Antibody Therapeutics international conferences, and the 2010 Annual Meeting of The Antibody Society, organized by IBC Life Sciences with contributions from The Antibody Society and two Scientific Advisory Boards, were held December...
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PMID: 21304271
PDF is available here.
PDF is available here.
Abstract
We describe a methodology to heterologously express, purify, and conduct in vitro characterization of several metalloforms of the AHL lactonase from Bacillus thuringiensis (AiiA). These procedures should be applicable to similar enzymes and will facilitate the production of more useful quorum-quench...
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PMID: 21031319
PDF is available here.
PDF is available here.
Abstract
This chapter describes the use of glutathione S-transferase (GST) gene fusion proteins as a method for inducible, high-level protein expression and purification from bacterial cell lysates. The protein is expressed in a pGEX vector, with the GST moiety located at the N terminus followed by the targe...
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PMID: 20978970
PDF is available here.
PDF is available here.
Abstract
Protein fusion technology has enormously facilitated the efficient production and purification of individual recombinant proteins. The use of genetically engineered affinity and solubility-enhancing polypeptide "tags" has increased greatly in recent years and there now exists a considerable repertoi...
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PMID: 20978965
PDF is available here.
PDF is available here.
Abstract
Maltose-binding protein (MBP) is one of the most popular fusion partners being used for producing recombinant proteins in bacterial cells. MBP allows one to use a simple capture affinity step on amylose-agarose columns, resulting in a protein that is often 70-90% pure. In addition to protein-isolati...
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PMID: 20978971
PDF is available here.
PDF is available here.
Abstract
Expression of recombinant proteins in the baculovirus/insect cell expression system is employed because it enables post-translational protein modification and high yields of recombinant protein. The system is capable of facilitating the functional expression of many proteins - either secreted or int...
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PMID: 20978972
PDF is available here.
PDF is available here.
Abstract
This article defines protein stability, emphasizes its importance and surveys some notable recent publications (2004-2008) in the field of protein stability/stabilization. Knowledge of the factors stabilizing proteins has emerged from denaturation studies and from study of thermophilic (and other ex...
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PMID: 20978963
PDF is available here.
PDF is available here.
Abstract
I glutaredoxins, GrxC1 and C4, CGYC and CPYC, respectively, were transformed into CCMC or CCMS. All the recombinant mutated proteins bind [2Fe-2S] centers into holodimers, whereas monomeric apoforms possess glutathione-dependent reductase activity. The functionally important, hydrophobic GALWL C-ter...
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PMID: 21094149
PDF is available here.
PDF is available here.
Abstract
We sought to develop an active method to increase the circulation half-life of proteins by binding to erythrocytes in blood. Screening a naive phage-displayed peptide library against whole mouse erythrocytes yielded a 12 amino acid peptide (ERY1) that binds the erythrocyte surface with high specific...
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PMID: 20831257
PDF is available here.
PDF is available here.
Abstract
A DNA vaccine encoding the whole segment of the Derp2 allergen could prevent allergic airway inflammation in a Derp2 allergen-induced allergic airway inflammation mouse model.
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PMID: 21337914
PDF is available here.
PDF is available here.
Acta Biomaterialia (6)12 2010
Abstract
We have used peptide motifs that have been selected to bind to gold, platinum, glass and titanium to modify surfaces with poly(ethylene glycol) anti-fouling polymer and the integrin-binding RGD sequence. The peptides have several advantages over conventional molecular immobilization techniques; they...
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PMID: 20601249
PDF is available here.
PDF is available here.
Abstract
We report for the first time a soluble PQQ-glucose dehydrogenase that is twice more active than the wild type for glucose oxidation and was obtained by combining site directed mutagenesis, modelling and steady-state kinetics. The observed enhancement is attributed to a better interaction between the...
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PMID: 21036156
PDF is available here.
PDF is available here.
Abstract
We suggest that cyclic peptides are an underdiscovered class of molecules and that many more will be discovered in the near future. There is much interest in understanding the mechanism of cyclization of circular proteins and the role of the cyclic backbone in defining structure and activity. In thi...
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PMID: 20835453
PDF is available here.
PDF is available here.
Abstract
Abstract
β-Turns are secondary structure elements not only exposed on protein surfaces, but also frequently found to be buried in protein-protein interfaces. Protein engineering so far considered mainly the backbone-constraining properties of synthetic β-turn mimics as parts of surface-exposed loops. A β-...
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PMID: 20937907
PDF is available here.
PDF is available here.
Abstract
Antimicrobial peptides will be attractive and potential candidates as peptide drugs because of their efficient action against microbes and low toxicity to mammal cells. To improve their antibacterial activity, some modifications needs to be made. In this research, the hybrid peptide gene Attacin-Tha...
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PMID: 19967452
PDF is available here.
PDF is available here.
Current Drug Safety (5)4 2010
Abstract
Protein therapeutics offer distinct advantages over other classes of drugs largely due to the high level of target specificity and generally low toxicity. Problems have, however, been encountered with some protein therapeutics inducing undesirable immune responses in patients. This i...
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PMID: 20615174
PDF is available here.
PDF is available here.
Abstract
These results are in full agreement with previously described affinity-labeling reactions performed with GABA(A) noncompetitive blockers (Perret et al. J. Biol. Chem.1999, 274, 25350-25354)....
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PMID: 20839772
PDF is available here.
PDF is available here.
Abstract
Bacteria are particularly convenient for producing recombinant proteins for purification purposes. To monitor induction as well as the levels of recombinant protein expression, it is important to have a rapid, simple method for estimating bacterial protein expression. This protocol describes the pre...
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PMID: 20810630
PDF is available here.
PDF is available here.
Abstract
Bacteria are particularly convenient for producing recombinant proteins for purification purposes. Suitable extraction methods for bacterial cells include sonication, glass bead milling, grinding with alumina or sand, high-pressure shearing using the French pressure cell (French Press), and lysozyme...
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PMID: 20810631
PDF is available here.
PDF is available here.
Abstract
The high levels of expression produced by molecular cloning techniques make bacteria particularly useful for producing recombinant proteins. However, these proteins often are difficult to purify, owing to their tendency to aggregate and precipitate within the bacteria as insoluble inclusion bodies....
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PMID: 20810632
PDF is available here.
PDF is available here.
Abstract
We describe an alternative strategy using stable transfection pool technology. Using stable transfection pools, gram quantities of monoclonal antibody (Mab) can be generated within 2 months post-transfection. Expression levels for monoclonal antibodies can be achieved ranging from 100 mg/L to over 1...
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PMID: 20564356
PDF is available here.
PDF is available here.
Abstract
We successfully isolated approximately 24 mg/L 3-O-rhamnosyl quercetin and 12.9 mg/L 3-O-rhamnosyl kaempferol upon feeding of 0.2 mM of the respective flavonoids and were also able to isolate 3-O-allosyl quercetin. Thus, this study reveals a method that might be useful for the biosynthesis of rhamno...
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PMID: 20506539
PDF is available here.
PDF is available here.
Abstract
We investigated the genomic mutation rates of the hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT) gene using a 6-thioguanine (6-TG) assay under various concentrations of MTX selection in CHO cells. Our results show that the 6-TG resistance increased as the MTX concentration increased...
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PMID: 20506532
PDF is available here.
PDF is available here.
Abstract
A recombinant monoclonal antibody produced by Chinese hamster ovary (CHO) cell fed-batch culture was found to have amino acid sequence misincorporation upon analysis by intact mass and peptide mapping mass spectrometry. A detailed analysis revealed multiple sites for asparagine were being randomly s...
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PMID: 20506364
PDF is available here.
PDF is available here.
Abstract
I transmembrane FcgammaRs were expressed in Escherichia coli and their interactions to subclass IgGs (IgG1, IgG2, IgG3, and IgG4) antibodies were analyzed. Expression using fully synthetic E. coli codon optimized FcgammaR genes and optimization of sequences for N-terminal translation initiation regi...
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PMID: 20506277
PDF is available here.
PDF is available here.
Abstract
We have developed an orthogonal protease to selectively activate each isoform in human cells. Our approach uses a split-tobacco etch virus (TEV) protease under small-molecule control, which we call the SNIPer, with caspase alleles containing genetically encoded TEV cleavage sites. These studies reve...
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PMID: 20723762
PDF is available here.
PDF is available here.
Abstract
We report improved production and purification protocols that have allowed detailed chemical analyses of these compounds. The polyketide synthase product 8-deoxy-16-descarboxyl-16-methyl-amphoteronolide B was identified for the first time. In addition, the ketoreductase 10 domain of the polyketide s...
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PMID: 20571619
PDF is available here.
PDF is available here.
Abstract
We mutated positions 158 and 160 in several conventional red-shifted FPs, including mNeptune, mCherry, mStrawberry, mOrange, and mKO, and the resulting FP variants exhibited LSS fluorescence emission in a wide range of wavelengths from 560 to 640 nm. These data suggest that different chromophores fo...
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PMID: 20681709
PDF is available here.
PDF is available here.
Abstract
Abstract
Viability decreased by 31% +/- 2% for Panc-1 cells (P < .0001) and 34% +/- 1% (P < .0001) for Hep 3B cells after treatment. Both cell lines demonstrated a 4-fold increase activated caspase-3 expression after high dose treatment (P < .01), and 5-fold increase in ASS-1 expression (P < .002). Ki-67 exp...
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PMID: 20466402
PDF is available here.
PDF is available here.
Abstract
Recombinant protein expression in Escherichia coli (E. coli) is simple, fast, inexpensive, and robust, with the expressed protein comprising up to 50 percent of the total cellular protein. However, it also has disadvantages. For example, the rapidity of bacterial protein expression often results in...
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PMID: 20814932
PDF is available here.
PDF is available here.
Abstract
gamma-Lactamase belongs to the amidase. The (+) gamma-lactamses can be applied in the kinetic resolution of racemic gamma-lactam, which can produce (-) gamma-lactam efficiently. The (-) gamma-lactam enantiomer is an important synthon for the synthesis of carbocyclic nucleosides. Up to now seven stra...
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PMID: 20931864
PDF is available here.
PDF is available here.
Abstract
We have established a strategy to produce and purify recombinant farnesylated guanylate-binding protein 1 (hGBP1), a dynamin-related large GTPase. Our system is based on the coexpression of hGBP1 with the two subunits of human farnesyltransferase in Escherichia coli and a chromatographic separation...
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PMID: 20348589
PDF is available here.
PDF is available here.
Abstract
These results suggest that our method will be useful in obtaining a large quantity of mPEG(10K)-cGLP-1 for further study and mPEG(10K)-cGLP-1 might find a role in the therapy of type 2 diabetes through C-terminal site-specific PEGylation....
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PMID: 19728172
PDF is available here.
PDF is available here.
Abstract
The recombinant DNA pLR5cat_PSAB in which pediocin PA-1 structural and immunity genes (pedAB) fused with the promoter and deduced signal sequence of an alpha-amylase gene from a bifidobacterial strain were inserted in pLR5cat, an Escherichia coli-lactobacilli shuttle vector was transferred to Lactob...
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PMID: 20798585
PDF is available here.
PDF is available here.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (26)8 2010
Abstract
To investigate the immune responses and the protection induced by the transgenic Alfalfa (Medicago sativa) containing Eg95-EgA31 fusion gene of Echinococcus granulosus against Eg protoscoleces.
The leaf protein was extracted from the transgenic alfalfa by heat-coagulation method, its concentration w...
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PMID: 20619100
PDF is available here.
PDF is available here.
Bioresource Technology (101)15 2010
Abstract
Polyhydroxyalkanoate synthase gene phaC(ah) in Aeromonas hydrophila strain 4AK4 was deleted and its function was replaced by phaC1(ps) cloned from Pseudomonas stutzeri strain 1317 which favors 3-hydroxyhexanoate (3HHx) and longer chain length monomers. Genes fadD and fadL encoding Escherichia coli a...
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PMID: 20236821
PDF is available here.
PDF is available here.
Abstract
I (Fe(+4)=O, porph+(.)) or Compound ES (Fe(+4)=O, Trp+(.)). R48A/W191F formed a Fe(+2)O2 complex upon photolysis of Fe(+2)CO in the presence of O2, and N-hydroxyguanidine was observed to react with this species to produce products, distinct from N-nitrosoguanidine, that gave a positive Griess reacti...
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PMID: 20346907
PDF is available here.
PDF is available here.
Journal of Biotechnology (148)4 2010
Abstract
Amarantin is the predominant seed storage protein from amaranth. It shows a high content of essential amino acids, making this protein important from a nutritional viewpoint. The protein has two disulfide linked subunits: acidic and basic. Acidic subunit has the potential as a functional and nutrace...
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PMID: 20561545
PDF is available here.
PDF is available here.
Abstract
Abstract
We report an efficient biocatalytic process to replace a recently implemented rhodium-catalyzed asymmetric enamine hydrogenation for the large-scale manufacture of the antidiabetic compound sitagliptin. Starting from an enzyme that had the catalytic machinery to perform the desired chemistry but lac...
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PMID: 20558668
PDF is available here.
PDF is available here.
Abstract
We describe the de novo computational design and experimental characterization of enzymes catalyzing a bimolecular Diels-Alder reaction with high stereoselectivity and substrate specificity. X-ray crystallography confirms that the structure matches the design for the most active of the enzymes, and...
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PMID: 20647463
PDF is available here.
PDF is available here.
Abstract
A simple biosensing strategy for the diagnosis of patients with hepatitis B virus (HBV) was developed. This study can be divided into two themes, both of which utilized gold-binding polypeptide (GBP) fusion proteins: HBV surface antigen PreS2 (HBsAg) detection with GBP-fused single chain antibody (G...
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PMID: 20602973
PDF is available here.
PDF is available here.