Synthesis of biotin–AMP conjugate for 5′ biotin labeling of RNA …
Nov 6, 2008 · In this protocol, we describe a simple procedure for 5′ RNA biotin labeling by one-step in vitro transcription. A biotin–AMP (adenosine 5′-monophosphate) conjugate, biotin-HDAAMP (adenosine...
Proximity labeling in mammalian cells with TurboID and split …
Nov 2, 2020 · TurboID is an engineered biotin ligase that uses ATP to convert biotin into biotin–AMP, a reactive intermediate that covalently labels proximal proteins.
活细胞内亚细胞结构蛋白质组学研究新技术——几种邻近标记策略 …
BPL标记其靶蛋白过程一般包含两个步骤：a. 催化生物素和ATP反应生成活化的生物素biotinoyl-5'-AMP，并将其结合在自身的活性位点上；b. 将活化的生物素共价偶联到与其结合的靶蛋白的赖氨酸ε-氨基上. a. Biotin + ATP ⇋ Bio-5'-AMP + PPi. b. Bio-5'-AMP + apo-Protein → Biotinoyl ...
A biotin targeting chimera (BioTAC) system to map small …
Dec 4, 2023 · The BioTAC system uses a universal recruitable biotin ligase chimera (miniTurboFKBP12 F36V) expression construct, facilitating rapid application to any system of interest without the need for ...
DNA Modifications Enabling Proximity Biotinylation
Mar 8, 2023 · This protocol describes the use of TurboID and split-TurboID in proximity labeling applications for mapping protein-protein interactions and subcellular proteomes in live mammalian cells. TurboID is an engineered biotin ligase that uses ATP to convert biotin into biotin-AMP, a reactive intermediate that covalently labels proximal proteins.
Molecular Spatiomics by Proximity Labeling
May 5, 2022 · These enzymes generate biotin-AMP ester (biotin-AMP) through a coupling reaction between biotin and ATP within the enzyme active site. Since biotin-AMP is an electrophilic activated ester, it reacts with lysine residues on …
Molecular Spatiomics by Proximity Labeling - PMC - National …
May 5, 2022 · These enzymes generate biotin-AMP ester (biotin-AMP) through a coupling reaction between biotin and ATP within the enzyme active site. Since biotin-AMP is an electrophilic activated ester, it reacts with lysine residues on …
Proximity labeling in mammalian cells with TurboID and split ... - PubMed
TurboID is an engineered biotin ligase that uses ATP to convert biotin into biotin-AMP, a reactive intermediate that covalently labels proximal proteins. Optimized using directed evolution, TurboID has substantially higher activity than previously described biotin ligase-related proximity labeling methods, such as BioID, enabling higher ...
BirA*-protein A fusion protein based BioEnhancer amplifies
The R118G mutation attenuated the biotin-AMP binding affinity of the bacterial biotin ligase BirA*, offering a proximity-dependent labeling ability, which could be used as a signal amplifier. We built a BirA*-protein A fusion protein (BioEnhancer) that specifically binds to IgG and adds biotin tags to its proximal amine groups, enhancing the ...
Synthesis of biotin-AMP conjugate for 5' biotin labeling of RNA …
A biotin-AMP (adenosine 5'-monophosphate) conjugate, biotin-HDAAMP (adenosine 5'-(6-aminohexyl) phosphoramide; where HDA is 1,6-hexanediamine), is chemically synthesized. Transcription initiation by biotin-HDAAMP under the T7 phi 2.5 promoter produces 5' biotin-labeled RNA with high labeling efficiency.