The Experts below are selected from a list of 3594 Experts worldwide ranked by ideXlab platform
Ting Wang - One of the best experts on this subject based on the ideXlab platform.
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Expression, purification and identification of CtCVNH, a novel anti-HIV (Human Immunodeficiency Virus) protein from Ceratopteris thalictroides.
International Journal of Molecular Sciences, 2013Co-Authors: Yingjuan Su, Ting WangAbstract:CVN (cyanovirin-N) is an anti-HIV protein. CVNH (cyanovirin-N homology) represents its homology. In a previous study, we first reported the full-length sequences of the CVNH gene cloned from Ceratopteris thalictroides. Based on the finding, the coding sequence of CtCVNH was optimized in the study, and then a pET prokaryotic expression vector was constructed. The purification and identification of CtCVNH protein were investigated, as well. SDS-PAGE analysis indicated that a 31 kDa protein was overexpressed and mainly accumulated in the soluble fraction. Only a single protein was obtained after the Ni- Nitrilotriacetic Acid (NTA) affinity chromatography. The purified protein was identified to be the recombinant CtCVNH by both Western blot and peptide mass fingerprinting analysis.
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Expression, Purification and Identification of CtCVNH, a Novel Anti-HIV (Human Immunodeficiency Virus) Protein from
2013Co-Authors: Ceratopteris Thalictroides, Junbo Sun, Ting WangAbstract:Abstract: CVN (cyanovirin-N) is an anti-HIV protein. CVNH (cyanovirin-N homology) represents its homology. In a previous study, we first reported the full-length sequences of the CVNH gene cloned from Ceratopteris thalictroides. Based on the finding, the coding sequence of CtCVNH was optimized in the study, and then a pET prokaryotic expression vector was constructed. The purification and identification of CtCVNH protein were investigated, as well. SDS-PAGE analysis indicated that a 31 kDa protein was overexpressed and mainly accumulated in the soluble fraction. Only a single protein was obtained after the Ni- Nitrilotriacetic Acid (NTA) affinity chromatography. The purified protein was identified to be the recombinant CtCVNH by both Western blot and peptide mass fingerprinting analysis
Jacob Piehler - One of the best experts on this subject based on the ideXlab platform.
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maleimide photolithography for single molecule protein protein interaction analysis in micropatterns
Analytical Chemistry, 2011Co-Authors: Sharon Waichman, Oliver Beutel, Maniraj Bhagawati, Changjiang You, Jacob PiehlerAbstract:Spatial organization of proteins into microscopic structures has important applications in fundamental and applied research. Preserving the function of proteins in such microstructures requires generic methods for site-specific capturing through affinity handles. Here, we present a versatile bottom-up surface micropatterning approach based on surface functionalization with maleimides, which selectively react with organic thiols. Upon UV irradiation through a photomask, the functionality of illuminated maleimide groups was efficiently destroyed. Remaining maleimides in nonilluminated regions were further reacted with different thiol-functionalized groups for site-specific protein immobilization under physiological conditions. Highly selective immobilization of His-tagged proteins into tris(Nitrilotriacetic Acid) functionalized microstructures with very high contrast was possible even by direct capturing of proteins from crude cell lysates. Moreover, we employed phosphopantetheinyl transfer from surface-imm...
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four color single molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes
BioTechniques, 2010Co-Authors: Vanessa Derocco, Jacob Piehler, Trevor Anderson, Dorothy A Erie, Keith WeningerAbstract:To enable studies of conformational changes within multimolecular complexes, we present a simultaneous, four-color single molecule fluorescence methodology implemented with total internal reflection illumination and camera-based, wide-field detection. We further demonstrate labeling histidine-tagged proteins noncovalently with Tris-Nitrilotriacetic Acid (Tris-NTA)-conjugated dyes to achieve single molecule detection. We combine these methods to colocalize the mismatch repair protein MutSα on DNA while monitoring MutSα-induced DNA bending using Forster resonance energy transfer (FRET) and to monitor assembly of membrane-tethered SNARE protein complexes.
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high affinity labeling and tracking of individual histidine tagged proteins in live cells using ni2 tris Nitrilotriacetic Acid quantum dot conjugates
Nano Letters, 2009Co-Authors: Victor Roullier, Jacob Piehler, Samuel Clarke, Fabien Pinaud, Geraldine Gouzer, Dirk Schaible, Valerie Marchiartzner, Maxime DahanAbstract:Investigation of many cellular processes using fluorescent quantum dots (QDs) is hindered by the nontrivial requirements for QD surface functionalization and targeting. To address these challenges, we designed, characterized and applied QD−trisNTA, which integrates tris-Nitrilotriacetic Acid, a small and high-affinity recognition unit for the ubiquitous polyhistidine protein tag. Using QD−trisNTA, we demonstrate two-color QD tracking of the type-1 interferon receptor subunits in live cells, potentially enabling direct visualization of protein−protein interactions at the single molecule level.
Hyun-jong Paik - One of the best experts on this subject based on the ideXlab platform.
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encapsulation of nanoparticles using Nitrilotriacetic Acid end functionalized polystyrenes and their application for the separation of proteins
Advanced Functional Materials, 2012Co-Authors: Mohammad Abdul Kadir, Sujeong Kim, Hong Y Cho, Bongsoo Kim, Donghyeuk Choi, Sungu Lee, Bog G Kim, Sangwook Kim, Hyun-jong PaikAbstract:The use of Nitrilotriacetic Acid end-functionalized polystyrenes (NTA-PS) as a multifunctional nanocarrier for the aqueous dispersion of CdSe, γ-Fe2O3 and gold nanoparticles (NPs) is described. When the amphiphilic end- functionalized polystyrenes and NPs are dissolved together in tetrahydrofuran, the addition of water causes the spontaneous formation of micellar aggregates, resulting in the successful encapsulation and aqueous dispersion of NPs. Transmission electron microscopy (TEM), scanning electron microscopy (SEM), photoluminescence (PL) spectroscopy, and vibrating sample magnetometer (VSM) are used to characterize the structure and properties of the NPs-containing micellar aggregates (nanocarrier). After complexation of Ni2+ with NTA on the surface of the nanocarrier containing γ-Fe2O3, specific binding between Ni-NTA complex and histidine-tagged (His-tagged) proteins enables selective separation of His-tagged proteins using a magnet.
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Synthesis of Well-Defined (Nitrilotriacetic Acid)-End-Functionalized Polystyrenes and Their Bioconjugation with Histidine-Tagged Green Fluorescent Proteins
Macromolecules, 2011Co-Authors: Mohammad Abdul Kadir, Soundrarajan Nagasundarapandian, Sung Bo Ko, Hyun-jong PaikAbstract:We have synthesized Nitrilotriacetic Acid end-functionalized polystyrene (NTA–PS) for the controlled bioconjugation with histidine-tagged green fluorescence proteins (His6–GFP). NTA–PS was prepared using initiators containing tert-butyl protected NTA moiety via atom transfer radical polymerization (ATRP) of styrene; the protected tert-butyl group was subsequently removed at the α-chain end of polystyrene. The structure of NTA–PS was characterized using 1H NMR, 13C NMR, and GPC. NTA chain ends of the polystyrenes were complexed with Ni2+ to produce Ni–NTA–PS, of which the specific binding properties were studied by forming spherical aggregates with His6–GFP in aqueous phase. The reversible association of His6–GFP with polystyrene spherical aggregates (with Ni2+) was controlled with imidazole and monitored with fluorescence microscope. Again, Ni–NTA–PS produced well-defined micelles with His6–GFP in water/DMF (DMF 4 vol %) and the size of micelles decreased when excess imidazole was added.
Xixiang Zhang - One of the best experts on this subject based on the ideXlab platform.
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dopamine as a robust anchor to immobilize functional molecules on the iron oxide shell of magnetic nanoparticles
Journal of the American Chemical Society, 2004Co-Authors: Chenjie Xu, Keming Xu, Hongwei Gu, Rongkun Zheng, Xixiang Zhang, Bing XuAbstract:We report on the use of dopamine (DA) as a robust molecular anchor to link functional molecules to the iron oxide shell of magnetic nanoparticles. Using Nitrilotriacetic Acid (NTA) as the functional molecule, we created a system with an M/Fe2O3−DA−NTA (M = Co or SmCo5.2) nanostructure, which possesses high stability and specificity for separating histidine-tagged proteins. The well-established biocompatibility of iron oxide and the robust covalent bonds between DA and Fe2O3 render this strategy attractive for constructing biofunctional magnetic nanoparticles containing iron oxide.
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Nitrilotriacetic Acid modified magnetic nanoparticles as a general agent to bind histidine tagged proteins
Journal of the American Chemical Society, 2004Co-Authors: Xiaofen Zhong, Rongkun Zheng, Zhihong Guo, Xixiang ZhangAbstract:Using Nα,Nα-bis(carboxymethyl)lysine to react with FePt magnetic nanoparticles, we synthesized the FePt−NTA conjugate, which immobilizes Ni2+ ions and selectively binds to histidine-tagged proteins...
Yuchie Chen - One of the best experts on this subject based on the ideXlab platform.
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Nitrilotriacetic Acid coated magnetic nanoparticles as affinity probes for enrichment of histidine tagged proteins and phosphorylated peptides
Analytical Chemistry, 2007Co-Authors: Ya Shiuan Lin, Pei Jane Tsai, Cheng Tai Chen, Wei Yu Chen, Yuchie ChenAbstract:We herein demonstrate superparamagnetic Fe3O4 nanoparticles coated with Nitrilotriacetic Acid derivative (NTA) that can bind with different immobilized metal ions are capable of probing diverse target species. Immobilized Ni(II) on the surfaces of the NTA−magnetic nanoparticles have the capability of selectively trapping histidine (His)-tagged proteins such as a mutated streptopain tagged with 6× His, i.e., C192S (MW ∼42 kDa), from cell lysates. Enrichment was achieved by vigorously mixing the sample solution and the nanoparticles by pipetting in and out of a sample vial for only 30 s. After enrichment, the probe−target species could be readily isolated by magnetic separation. We also characterized the proteins enriched on the affinity probes using on-probe tryptic digestion under microwave irradiation for only 2 min, followed by matrix-assisted laser desorption/ionization mass spectrometry analysis. Using this enrichment and tryptic digestion, the target species can be rapidly enriched and characterized,...
