Abstract:
:Current advancements in biological protein discovery utilize bi-partite methods of fluorescence detection where chromophore and scaffold are uncoupled. One such technology, called fluorogen-activating proteins (FAPs), consists of single-chain-variable-fragments (scFvs) selected against small organic molecules (fluorogens) that are non-fluorescent in solution, but highly fluorescent when bound to the scFv. In unusual circumstances a scFv may activate similar fluorogens from a single chemical family. In this report we identified a scFv biosensor with fluorescence activity against multiple fluorogens from two structurally dissimilar families. In-vitro analysis revealed highly selective scFv-ligand interactions at sub-micromolar ranges. Additionally, each scFv-fluorogen complex possesses unique excitation and emission spectra, which allows broader detection limits from the biosensor. Further analysis indicated that ligand activation, regardless of chemical family, occurs at a common scFv binding region that proves flexible, yet selective for fluorogen binding. As a protein reporter at the surface of mammalian cells, the scFv revealed bright signal detection and minimal background. Additionally, when tagged to a G-protein-coupled receptor, we observed agonist dependent signaling leading to protein traffic from cell surface to endosomes via multi-color fluorescence tracking. In summary, this report unveils a noncanonical scFv biosensor with properties of high ligand affinity and multi-channel fluorescence detection, which consequently offers expanded opportunities for cellular protein discovery.
journal_name
Protein Pept Lettjournal_title
Protein and peptide lettersauthors
Gallo E,Wienbar S,Snyder AC,Vasilev KV,Armitage BA,Jarvik JWsubject
Has Abstractpub_date
2014-01-01 00:00:00pages
1289-94issue
12eissn
0929-8665issn
1875-5305pii
PPL-EPUB-60972journal_volume
21pub_type
杂志文章abstract::Porins are integral membrane proteins found in the outer membrane of bacteria, mitochondria and chloroplasts. Herein, we have reviewed sequence and structural understanding about bacterial porins. The first porin structure from Rhodobacter capsulatus at 1.8 Å resolution in 1991 till the recent structural advancement, ...
journal_title:Protein and peptide letters
pub_type: 杂志文章
doi:10.2174/0929866521666140827110755
更新日期:2015-01-01 00:00:00
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journal_title:Protein and peptide letters
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doi:10.2174/0929866523666160223122415
更新日期:2016-01-01 00:00:00
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journal_title:Protein and peptide letters
pub_type: 杂志文章,评审
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更新日期:2004-08-01 00:00:00
abstract::3-Isopropylmalate dehydrogenase (IPMDH) from Mycobacterium tuberculosis (Mtb) may be a target for specific drugs against this pathogenic bacterium. We have expressed and purified Mtb IPMDH and determined its physicalchemical and enzymological properties. Size-exclusion chromatography and dynamic light scattering measu...
journal_title:Protein and peptide letters
pub_type: 杂志文章
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更新日期:2014-01-01 00:00:00
abstract:BACKGROUND:Wheat germ, one of the byproducts of flour milling, contains abundant physiologically active components. Globulins in wheat germ are a class of high-quality functional proteins and have received widespread attention. However, the composition of wheat germ globulin( WGG)and the structure of the typical protei...
journal_title:Protein and peptide letters
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doi:10.2174/0929866524666171026123054
更新日期:2018-02-08 00:00:00
abstract::We report the physical-chemical characterization of the major alcohol-soluble proteins present in seeds of pearl millet (Pennisetum glaucum) by SDS-PAGE, bidimensional gel electrophoresis, MALDI-TOF/MS and RP-HPLC. We demonstrate the presence of three major prolamins, called A-, B- and C-pennisetin with mass values ar...
journal_title:Protein and peptide letters
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doi:10.2174/0929866023408724
更新日期:2002-06-01 00:00:00
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journal_title:Protein and peptide letters
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更新日期:2008-01-01 00:00:00
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journal_title:Protein and peptide letters
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doi:10.2174/092986608784966958
更新日期:2008-01-01 00:00:00
abstract::Saturation Transfer Difference NMR (STD NMR) is used for the detection of the binding constant of the decapeptide RRRDDDSDDD with CK2α, the catalytic subunit of the proteinkinase CK2. For this work a valid irradiation frequency of the CK2α had to be found ensuring that no peptide resonance is affected by the irradiati...
journal_title:Protein and peptide letters
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更新日期:2012-09-01 00:00:00
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
pub_type: 杂志文章,评审
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更新日期:2009-01-01 00:00:00
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journal_title:Protein and peptide letters
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doi:10.2174/0929866526666191204124245
更新日期:2020-01-01 00:00:00
abstract:BACKGROUND:Cysteine S-sulfenylation is a major type of dynamic post-translational modification of the protein that plays an important role in regulating many biological processes in both of prokaryotic and eukaryotic species. To understand the function of S-sulfenylated proteins, identification of S-sulfenylation sites...
journal_title:Protein and peptide letters
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更新日期:2018-01-01 00:00:00
abstract::The present paper reviews the use of expressed protein ligation for the biosynthesis of backbone cyclized polypeptides. This general method allows the in vivo and in vitro biosynthesis of cyclic polypeptides using recombinant DNA expression techniques. Biosynthetic access to backbone cyclic peptides opens the possibil...
journal_title:Protein and peptide letters
pub_type: 杂志文章,评审
doi:10.2174/0929866054864274
更新日期:2005-11-01 00:00:00
abstract:BACKGROUND:Haloalkane dehalogenases (EC 3.8.1.5, HLDs) are α/β-hydrolases which catalyze the irreversible cleavage of carbon-halogen bonds of haloalkanes, producing an alcohol, a halide and a hydrogen ion. Haloalkanes are acutely toxic to animals and humans and their toxic effects are mainly observed in the liver, kidn...
journal_title:Protein and peptide letters
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doi:10.2174/0929866524666170621094531
更新日期:2017-01-01 00:00:00
abstract::Chemical probing of histidine residues using specific modifiers, iodoacetic acid (IAA) and diethylpyrocarbonate (DEP) resulted in the inactivation of phytase (phy A). The kinetic theory of the substrate reaction during the modification of enzyme activity was applied to a study of the kinetics of the course of inactiva...
journal_title:Protein and peptide letters
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doi:10.2174/092986606777145788
更新日期:2006-01-01 00:00:00
abstract::Although macromolecular crystallography is a widely supported technique at synchrotron radiation facilities throughout the world, there are, in comparison, only very few beamlines dedicated to small-molecule crystallography. This limited provision is despite the increasing demand for beamtime from the chemical crystal...
journal_title:Protein and peptide letters
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doi:10.2174/0929866523666160115131217
更新日期:2016-01-01 00:00:00
abstract::Natural, synthetic and analogues of peptides have shown prospects for application in cancer chemotherapy. Notably, some food protein-derived peptides are known to possess anticancer activities in cultured cancer cells, and also in animal cancer models via different mechanisms including induction of apoptosis, cell cyc...
journal_title:Protein and peptide letters
pub_type: 杂志文章,评审
doi:10.2174/0929866523666160816152755
更新日期:2017-01-01 00:00:00
abstract::Transgelin was solubly expressed in E. coli. Crystals of transgelin have been grown at 291K using sodium formate or PEG-4000 as precipitants. X-ray diffraction by the crystal extends to 2.3 A resolution. The crystal belongs to the space group P2(1), with the unit cell parameters a=39.3, b=61.9, c=56.0 A and beta=90.2 ...
journal_title:Protein and peptide letters
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更新日期:2007-01-01 00:00:00
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journal_title:Protein and peptide letters
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abstract::Catalase is an enzyme that occurs in almost all aerobic organisms. Its main metabolic function is to prevent oxidative damage to tissues induced by hydrogen peroxide which is a strong oxidizing agent. Catalase is very effective in performing this task, since it has the highest turnover rate among all the enzymes. The ...
journal_title:Protein and peptide letters
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更新日期:2011-09-01 00:00:00
abstract::Like those in mammals, heterotrimeric G protein complexes have been implicated in signal transduction pathways in plants; however, the subunits themselves have not been isolated. In this study, the rice heterotrimeric G protein subunits α (Gα) and β (Gβ) were purified by affinity chromatography using anti-Gα and -Gβ a...
journal_title:Protein and peptide letters
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更新日期:2012-03-01 00:00:00
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journal_title:Protein and peptide letters
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更新日期:2015-01-01 00:00:00
abstract::Apelin, an endogenous ligand of the G-protein-coupled receptor APJ, is a novel myokine and may play a key role in regulating energy metabolism. The purpose of the present study was to investigate the effects of hypoxic exposure, exercise, and hypoxic exercise training on the expression of apelin and APJ in skeletal mu...
journal_title:Protein and peptide letters
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更新日期:2017-01-01 00:00:00
abstract::Heparin is naturally occurring polysaccharides which interacts with seminal plasma proteins and regulate multiple steps in fertilization process. Qualitative and quantitative information regarding the affinity for heparin-seminal plasma proteins interactions is not generally well documented and there are no reports of...
journal_title:Protein and peptide letters
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更新日期:2012-08-01 00:00:00
abstract::The small heat shock protein SsHSP14.1 from the hyper-thermophilic archeaon, Sulfolobus solfataricus (S. solfataricus) was able to protect proteins from thermal aggregation and prevent enzymes from heat induced inactivation. According to the 3D (dimensional) structural model of SsHSP14.1 developed by us before, the re...
journal_title:Protein and peptide letters
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更新日期:2011-03-01 00:00:00
abstract::Calreticulin is a soluble endoplasmic reticulum chaperone, which has a relatively low melting point due to its remarkable structure with a relatively high content of flexible structural elements. Using far ultraviolet circular dichroism (CD) spectroscopy and a fluorescent dye binding thermal shift assay, we have inves...
journal_title:Protein and peptide letters
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更新日期:2013-05-01 00:00:00
abstract::The Saccaromices cerevisiae D-serine dehydratase is a pyridoxal 5'-phosphate dependent enzyme that requires zinc for its function. It catalyses the conversion of D-serine into pyruvate and ammonia with the K(m) and k(cat) values of 0.39 mM and 13.1 s(-1) respectively. In this work, a new methodology for monitoring D-s...
journal_title:Protein and peptide letters
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doi:10.2174/092986612800494002
更新日期:2012-06-01 00:00:00
abstract:BACKGROUND:It is currently believed that protein folding rates are influenced by protein structure, environment and temperature, amino acid sequence and so on. We have been working for long to determine whether and in what ways mRNA affects the protein folding rate. A large number of palindromes aroused our attention i...
journal_title:Protein and peptide letters
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更新日期:2020-01-01 00:00:00