biotinylation

..of naturally biotinylated substrates are highly conserved across evolutionary boundaries, and cross-species biotinylation has been demonstrated in several systems...

..b>Biotinylation is an attractive approach for protein complex purification due to the very high affinity of avidin/..

..First, outer membrane proteins were enriched by biotinylation of intact cells using the reagent NHS (N-hydroxysuccinimide)-PEO(12) (polyethylene oxide)-biotin with ..

..We labeled the most exposed surface proteins using two impermeant biotinylation reagents that differed only in length...

..In dissecting the pluripotent state in mouse embryonic stem (mES) cells, we have used in vivo biotinylation of critical transcription factors for affinity purification of protein complexes and chromatin ..

..We also show that internalization of GluR1, as triggered by NMDAR activation, does not require SAP97. These results implicate GluR1-SAP97 interactions in mechanisms underlying AMPA receptor targeting...

..This suppressed proliferation of mesenchymal cells may delay or inhibit the growth of palatal processes in embryos and thus it may partially contribute to the mechanisms for cleft palate induction...

..Both strategies yielded efficient biotinylation of tagged HNE-protein conjugates, but click chemistry was found to be superior for the recovery of ..

..However, in vitro chemical techniques for protein biotinylation are not always successful, with some common problems being a lack of reaction specificity, inactivation of ..

In dissecting the pluripotent state in mouse embryonic stem (ES) cells, we have employed in vivo biotinylation of critical transcription factors for streptavidin affinity purification of protein complexes and constructed a protein-..

..Our data demonstrate that AMPH leads to an increase of NET in endosomes of single boutons and varicosities in a Rab11-dependent manner...

..Protein catch and photorelease shows that both HSA and ApoA1 in human plasma undergo significant modification by aHNE...

..the dynamic aspects underlying PcG composition and function we have established an inducible version of the biotinylation tagging approach to purify Polycomb and associated factors from Drosophila embryos...

..Since this method does not require specific antibodies and is inexpensive, sensitive, and reliable, it should be useful for detecting protein-protein interactions in cells...

..Site-specific biotinylation of proteins in vivo can be achieved by co-expressing in mammalian cells the protein of interest fused to a 15 ..

....

..Site-specific biotinylation and streptavidin staining take only a few minutes, while expression of BirA takes 4 d and expression of ..

..We employed in vivo perfusion of biotinylation reagent through rat brain microvessels to domain-specifically label proteins exposed on the microvessel ..

..Such a bias leads to protein-protein interactions when the lead monomer stalls owing to the presence of the streptavidin block...

..This review will examine the biotin-avidin technology for the study of cell surface proteins, discussing reagents and techniques as well as examples of applications in quantitative proteomics...

..These results indicate that the detection sensitivity, simplicity (single tube reaction) and speed (2 h) of this direct labeling protocol may be ideal for diagnostic applications that do not require target amplification...

..Recently, a team of investigators failed to detect biotinylated histones and proposed that biotinylation is not a natural modification of histones, but rather an assay artifact...

..e. biotinylation of histones...

..The detection limit of both methods was equivalent and corresponds to 1 amol of biotinylated DNA attached on an array. Scanning and data analysis of the array were obtained with a colorimetric-based workstation...

Using Biacore's new regenerateable streptavidin capture (CAP) sensor chips, we investigated a number of biotinylation conditions for target ligands...

..Here, we describe the results of a chemical proteomic study, based on the in vivo biotinylation of vascular proteins in lymphoma-bearing mice followed by mass spectrometric and bioinformatic analysis, to ..

..No significant detrimental effects on cell viability or death were observed following cell biotinylation. SPIO-labeled cells exhibited signal loss compared to non-SPIO-labeled cells by MRI in vitro...

....

..Therefore, this mouse can be crossed to any transgenic mouse to obtain efficient biotinylation of an Avi-tagged protein for the purpose of protein (complex) purification.

..Here, we present a method for cell surface protein biotinylation in the acute hippocampal slice, the most commonly used preparation for electrophysiological recordings...

....

....

..Secondly, we tested in vivo and in vitro site-directed biotinylation with two different tags, consisting of either 15 (AviTag) or 72 amino acids (BioEase tag), which serve as a ..

..All three apoE isoforms were equally detected in the nucleus. These studies provide new evidence that apoE may be targeted to the nucleus and shed light on factors that regulate this process...

..The wild type BirA protein is much less active in these reactions. The biotinylation reaction is proximity-dependent in that a greater extent of biotinylation was seen when the mutant ligase was ..

..g. acetylation, methylation, and biotinylation. We sought to identify amino-acid residues that are biotinylated in histone H4, and to determine whether ..

..BCCP becomes an active substrate for carboxylation and carboxyl transfer, after biotinylation of its canonical lysine residue by biotin protein ligase (BPL)...

..Histone biotinylation has been implicated in heterochromatin structures, DNA repair, and mitotic chromosome condensation...

..6mg) resulted in the labeling of erythrocytes released in blood circulation after the first biotinylation step, with a lower intensity of biotin...

..This new cell line has the potential for widespread application to envelope pseudotypes compatible with lentiviral vector production...

..Monovalent streptavidin should find general application in biomolecule labeling, single-particle tracking and nanotechnology...

A fluoride/amine-cleavable phosphoramidite designed for biotinylation, phosphorylation, and affinity purification of synthetic oligonucleotides was synthesized and coupled efficiently to the 5'-end of DNA on a solid-phase automatic ..

..The primary aim of this study was to identify biotinylation sites in histone H2A and its variant H2AX...

A fluoride-cleavable phosphoramidite for biotinylation was designed, synthesized and coupled efficiently to the 5'-end of DNA on an automatic synthesizer...

..Recently, we reported a novel modification of histones: biotinylation of lysine residues...

..Together, our data show that ubiquitination and endocytosis of DM-GRASP in concert regulate its cell surface concentration, which is crucial for its function in axon navigation...

..displayed on bacteriophage lambda heads to determine the minimal sequence length that is necessary to effect biotinylation in vivo...

..It has been shown that biotinylation of histone H4 at lysine-12 in histone H4 (K12Bio-H4) is associated with repression of a number of genes...

..Collectively, this study reveals a novel diet-dependent epigenetic mechanism that could affect cancer risk...

..structures provided evidence for small, localized conformational changes in the biotin-binding region upon biotinylation of the protein. These structural changes may be important for regulating specific protein-protein interactions...

..of intercellular contacts (BLINC), a method for imaging protein interactions based on interaction-dependent biotinylation of a peptide by E...

..b>Biotinylation is a relatively rare modification, with between one and five biotinylated protein species found in different ..

..These data indicate that a conformational change accompanies biotinylation of the biotin domain...

..T cells of identical specificity but in different activation states varied greatly in their ability to be detected by class II MHC oligomers...

..This study tested the hypothesis that BirA ligase catalyzes the biotinylation of eukaryotic histones...

..Here, we review recent findings that biotinylation of lysine-12 in histone H4 (H4K12bio) is an epigenetic mechanism to repress long terminal repeat (LTR) ..

Covalent modifications of histones play a role in regulating telomere attrition and cellular senescence. Biotinylation of lysine (K) residues in histones, mediated by holocarboxylase synthetase (HCS), is a novel diet-dependent mechanism ..

Biotin protein ligase (BPL) catalyzes the biotinylation of the biotin carboxyl carrier protein (BCCP) only at a special lysine residue. Here we report the first structure of BPL...

..Our findings suggest that hypoxia regulates both expression and activity of CA IX in order to enhance the extracellular acidification, which may have important implications for tumor progression...

..This behaviour coincides with the occurrence of an immobile band previously observed in electrophoretic experiments, indicating the formation of extremely large DNA-STV aggregate networks...

We present intein-mediated approaches for efficient biotinylation of proteins site-specifically...

..The vesosome is a better mimic of eukaryotic cell structure and demonstrates the benefits of multiple internal bilayer-enclosed compartments...

..We previously developed intein-mediated strategies for protein biotinylation and site-specific protein microarray generation...

..These results indicate that avidin has access to a number of sites of the DHPR within fully assembled (RyR1-containing) junctions, but not to the alpha1S C-terminal, which appears to be occluded by the presence of RyR1...

..The specificity and versatility of the labeling should allow it to become an important tool for studying and manipulating cell surface proteins and for complementing existing approaches in cell surface engineering...

Site-specific, enzymatic biotinylation of recombinant proteins can be exploited to circumvent many problems associated with the use of biotinylating reagents in vitro and to overcome some of their inherent limitations...

..b>Biotinylation studies revealed that modulation of ENaC function is due to the reduced apical expression of channel proteins...

..Thus, semiconductor nanocrystal fluorophores offer a more stable and quantitative mode of fluorescence in situ hybridization (FISH) for research and clinical applications...

..Using this approach we efficiently recovered and quantified RNAs bound to cytoplasmic poly(A)-binding protein (PABP) and to nuclear human transformer 2 (hTra-2) with minimal background...

..We describe a highly efficient system for in vivo biotinylation of transcription factors in the yeast Saccharomyces cerevisiae, which is based on the bacterial BirA biotin ..

..Tagged proteins were visualised through streptavidin probing of Western blots. Seven biotinylated proteins of Y. pestis were identified including two porins and the putative virulence factor catalase peroxidase...

..Results from cell line and patient sample evaluations indicate that this method may be easily incorporated into the diagnostic molecular laboratory as a rapid and cost-effective alternative to currently employed techniques...

b>Biotinylation in vivo is an extremely selective post-translational event where the enzyme biotin protein ligase (BPL) catalyzes the covalent attachment of biotin to one specific and conserved lysine residue of biotin-dependent enzymes...

....

To develop a novel procedure for isolating and characterizing cryptococcal cell-surface proteins using biotinylation, fluorescein isothiocyanate (FITC)-streptavidin, flow cytometry and associated ligand-receptor analysis, confocal ..

..prepared by reaction with d-biotinyl-6-aminocaproic acid N-hydroxysuccinimide ester and identified the biotinylation sites by trypsin digestion, reverse-phase chromatography, and electrospray mass spectrometry analyses...

b>Biotinylation of proteins is an attractive alternative to 'epitope-tagging', due to the strong biotin-(strept)avidin interaction and to the wide commercial availability of reagents for detection and purification of biotinylated ..

..Both fluorescein and a benzophenone photoaffinity probe are incorporated, with total labeling times as short as 20 min...

..The applicability of this kind of bioconjugation strategy to generate sensor-probes for gene detection assays was demonstrated...

..To study these interactions, we developed a simple and fast procedure for the biotinylation of carbohydrates based on reductive amination...

..These studies suggest that wild-type torsinA undergoes anterograde transport along microtubules mediated by kinesin and may act as a molecular chaperone regulating kinesin activity and/or cargo binding...

..In this study, we demonstrated that hLHR is capable of forming receptor complexes. Our findings may expand the possibility of a splice variant of hLHR specifically modulating the functional property of the wild-type hLHR...

..Pulse-chase radiolabeling and cell surface biotinylation experiments in the pituitary corticotroph cell line AtT20 showed that the mature 60/65 kDa form that resides ..

..Nogo-A, -B, and -C share a 180-amino acid C-terminal domain with two highly conserved hydrophobic stretches that could form a channel or transporter in the ER and/or on the cell surface...

..Thus selective association between TRPCs generate distinct store-operated channels. Diversity of store-operated channels might be related to the physiology of the different cell types...

..Our results suggest that Tat might function as a nuclear regulator of PP1 and that interaction of Tat with PP1 is critical for activation of HIV-1 transcription by Tat...

..Further characterization using surface biotinylation and flow cytometry assays indicated that pig Duox2-Q686X was present at the plasma membrane, whereas human ..

..Conformational changes in the protein affect the accessibility of the residues in extracellular loop 5 and provide further evidence of large-scale changes in the structure of NaDC-1 during the transport cycle...

..fluorescent protein, was analyzed in polarized LLC-PK1 cells by confocal microscopy and surface-specific biotinylation. Deletion of the N-glycosylation sites at N1, N3, N5, and N7 but not at N2, N4, and N6 significantly slowed ..

..Hetero-oligomers of wild-type kAE1 with recessive kAE1 S773P or G701D, in contrast to the dominant kAE1 R589H mutant, were delivered to the plasma membrane...

..These results indicate that ANKRA could modulate the excitability of neurons by binding directly to endogenous BKCa channel and altering its gating kinetics in a calcium-dependent manner...

..It is responsible for the capture of Kitl at coat protein complex II-coated ER exit sites, leading to subsequent cell surface transport under physiological conditions...

..Confocal microscopy and cell surface biotinylation indicated that all altered proteins were properly localized at the cell membrane...

..These results indicate that 16K expression inhibits beta(1) integrin surface expression and spreading on matrix by a novel mechanism that results in reduced levels of functional beta(1) integrin...

..Finally, crosslinking of cyclic BTNL1AD3 peptide to a leukemic T-cell membrane surface receptor, as examined using a biotin-avidin blot, indicated a molecular weight of approximately 34,400...

..in generation of major histocompatibility complex (MHC) tetramer relies on application of a key technique, biotinylation of MHC molecule specifically on a single lysine residue using the BirA enzyme...

..In addition, adhesion of SE to these model substrata can be controlled by the orientation of the protein...

..Furthermore, we provide evidence that TG2 is a major component of the IGFBP-3 kinase activity present on breast cancer cell membranes...

..Our results indicate that ligand-induced reversal of controlled TMD dimerization by the Notch extracellular domain is unlikely to underlie the regulatory mechanism of intramembranous cleavage...

..These results suggest that H-Ras present in membranes of the recycling endosome might be activating signal cascades essential for the dynamic and function of the organelle...

..Thus, interactions between CNR/Pcdhalpha and Pcdhgamma regulate their surface expression and contribute to the combinatorial diversity of cell recognition proteins in the brain...

..tropicalis on the FN molecule. In Specific Aim Three, we will use surface biotinylation and immunoprecipitation to characterize other surface proteins in C...

..and all nascent proteins destined for GPI anchorage, their reaction with biotinylated GPIs will result in the biotinylation of all GPI-anchored proteins...

..polarized epithelial cells, the fate of chimeric molecules determined by immunoflorescence and surface biotinylation. Individual residues in each subdomain of the TM4 necessary for a proper sorting of each molecule will then be ..

..hormonal effects, experiments are planned to study the plasma membrane distribution of the SERT using biotinylation studies;to examine the role of specific ER subtypes;and to study possible involvement of brain-derived ..

..The advantage of this biotinylation approach is that it allows the imaging of brain tumors with different modalities including Magnetic Resonance (..

..B) Surface biotinylation of BK channels in BAEC to determine whether BK channel activation involves the trafficking of BK channels from ..

..B) Surface biotinylation of BK channels in BAEC to determine whether BK channel activation involves the trafficking of BK channels from ..

..We will employ cell biological approaches including surface biotinylation, Western Blotting, immunofluorescece and immunoelectron microscopic localization in all animal and cell ..

..In addition, we will employ surface biotinylation, surface chemiluminescence, co-immunoprecipitation analysis, and macromolecular assembly assays, to verify ..

..Specific affinity biotinylation methods, which proved to be successful in the identification of mRPE65 as a tRE binding protein, will be ..

..abundance on the kinetics of restitution of cellular polarity will be explored using immunohistochemistry and biotinylation studies to track movement of displaced NaK ATPase following injury and during recovery...

..of targeting and insertion of native transporters will be assessed in hepatocytes using immunoblot and biotinylation, and targeting and insertion of fluorescently tagged transporters will be assessed in liver cell lines using ..

..such as transfection with constitutively active and dominant negative constructs, western blotting, and biotinylation assays, to investigate the actions of 5-HT1B receptors in hippocampal slices taken from control animals and ..

..a glucose transporter protein (SGTP4) that was definitely demonstrated by immunolocalization and surface biotinylation to be exposed at the host/parasite interface in the worm tegument...

..Epithelial isolation, cell surface biotinylation, and Western blot overlay methods will be applied to identify the glycosylated or non-glycosylated M cell ..

..responses after treating cells with agonists and by quantifying internalization of CaRs using a surface biotinylation assay...

..Functional biochemical techniques such biotinylation/immunoprecipitation will complement morphologic studied to investigate a role for membrane traffic...

..and functional expression of mEH on the hepatocyte cell surface will be investigated using antibody and biotinylation reagents, confocal fluorescence microscopy, glycosylation site insertion technology, co- immunoprecipitation, ..

..of the OX26 MAb and streptavidin (SA) is prepared in parallel with the carboxyl-directed pegylation and mono-biotinylation of recombinant human BDNF...

..Vesicle fusion machinery will be analyzed by a combination of biochemical (biotinylation, immunoprecipitation, affinity chromatography), immunocytochemical and genetic (siRNA knock-down of SNARE ..

..synthetase (HCS) mediates the binding of biotin to histones (DNA-binding proteins) H2A, H3, and H4, and that biotinylation of histones causes gene repression...

..synthetase (HCS) mediates the binding of biotin to histones (DNA-binding proteins) H2A, H3, and H4, and that biotinylation of histones causes gene repression...

..negatives of G protein receptor kinases (GRK), arrestins, and dynamin, confocal microscopic and cell surface biotinylation experiments will examine the process of CHO-MT2 receptor internalization...