Schneller, weiter, TurboID – Modulation einer übereifrigen Biotin-Ligase
نویسندگان
چکیده
Abstract Protein-protein interactions are key elements of intracellular signalling and metabolic pathways. These can be revealed with the help proximity ligation screens, prominently biotinylation screens. This approach has profited from recent development highly active biotin ligase TurboID, which however also led to problems toxicity related its high basal activity. We have established a simple protocol improve TurboID performance enhance protein functionality.
منابع مشابه
Overexpression of biotin synthase and biotin ligase is required for efficient generation of sulfur-35 labeled biotin in E. coli
BACKGROUND Biotin is an essential enzyme cofactor that acts as a CO2 carrier in carboxylation and decarboxylation reactions. The E. coli genome encodes a biosynthetic pathway that produces biotin from pimeloyl-CoA in four enzymatic steps. The final step, insertion of sulfur into desthiobiotin to form biotin, is catalyzed by the biotin synthase, BioB. A dedicated biotin ligase (BirA) catalyzes t...
متن کاملBiotin and fluorescent labeling of RNA using T4 RNA ligase.
Biotin, fluorescein, and tetramethylrhodamine derivatives of P1-(6-aminohex-1-yl)-P2-(5'-adenosine) pyrophosphate were synthesized and used as substrates with T4 RNA ligase. In the absence of ATP, the non-adenylyl portion of these substrates is transferred to the 3'-hydroxyl of an RNA acceptor to form a phosphodiester bond and the AMP portion is released. E. coli and D. melanogaster 5S RNA, yea...
متن کاملSelective inhibition of biotin protein ligase from Staphylococcus aureus.
There is a well documented need to replenish the antibiotic pipeline with new agents to combat the rise of drug resistant bacteria. One strategy to combat resistance is to discover new chemical classes immune to current resistance mechanisms that inhibit essential metabolic enzymes. Many of the obvious drug targets that have no homologous isozyme in the human host have now been investigated. Ba...
متن کاملBiotin Protein Ligase Is a Target for New Antibacterials
There is a desperate need for novel antibiotic classes to combat the rise of drug resistant pathogenic bacteria, such as Staphylococcus aureus. Inhibitors of the essential metabolic enzyme biotin protein ligase (BPL) represent a promising drug target for new antibacterials. Structural and biochemical studies on the BPL from S. aureus have paved the way for the design and development of new anti...
متن کاملAn improved smaller biotin ligase for BioID proximity labeling
The BioID method uses a promiscuous biotin ligase to detect protein-protein associations as well as proximate proteins in living cells. Here we report improvements to the BioID method centered on BioID2, a substantially smaller promiscuous biotin ligase. BioID2 enables more-selective targeting of fusion proteins, requires less biotin supplementation, and exhibits enhanced labeling of proximate ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Biospektrum
سال: 2023
ISSN: ['0947-0867', '1868-6249']
DOI: https://doi.org/10.1007/s12268-023-1943-6