Biotinylated Metabolite
Biotinylated Metabolite - Related Videos
09:13
Cell-surface Biotinylation Assay
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2023
A cell can regulate the amount of particular proteins on its cell membrane through endocytosis, following which cell surface proteins are effectively sequestered in the cytoplasm. Once within a cell, these surface proteins can be either destroyed or “recycled” back to the membrane. The cell surface biotinylation assay provides researchers with a way to study these phenomena. The technique makes use of a derivative of the small molecule biotin, which can label surface proteins and then be...
01:18
Detection of Human Fecal Metabolites Using a Bacterial Biosensor
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2026
Begin with a bacterial biosensor culture suspended in a medium.These engineered bacteria produce a transcription factor that, upon binding to a specific fecal metabolite, activates a promoter associated with a fluorescent reporter gene.To this culture, add a filtered human fecal sample containing metabolites, such as lactate, that are involved in host-microbiome interactions.Dilute this mixture with a fresh medium to maintain an optimal reaction volume.Incubate without shaking to mimic the...
11:32
Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
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Cited by 20 •
2017
Here, we detail the method of Sequencing of Psoralen crosslinked, Ligated, and Selected Hybrids (SPLASH), which enables genome-wide mapping of intramolecular and intermolecular RNA-RNA interactions in vivo. SPLASH can be applied to study RNA interactomes of organisms including yeast, bacteria and humans.
06:43
In Vivo Proximity Biotinylation for Protein Interaction Studies in Paramecium tetraurelia
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Cited by 1 •
2025
The protocol presents a method for in vivo covalent attachment of biotin to proteins based on their proximity to a biotin ligase fused to a protein of interest. This modification allows for a selective enrichment of the proteins using streptavidin beads as needed in protein interaction studies.
12:49
Purification of Transcripts and Metabolites from Drosophila Heads
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Cited by 12 •
2013
We describe here the procedures for the extraction and purification of mRNA and metabolites from Drosophila heads. We are applying these techniques to better understand the cellular perturbations underlying neuronal degeneration. These methodologies can be easily scaled and adapted for other "omic" projects.
FAQs
ATP first activates bicarbonate, creating a reactive carboxyl group that can be transferred to the ureido ring of biotin. This energy-dependent sequence prepares biotin to carry the carboxyl group between catalytic sites or onto a substrate. The coupling of ATP use and group transfer is central to understanding the reaction mechanism.
The ureido ring provides the site where the carboxyl group becomes attached to biotin during biotin-dependent carboxylation. Biotin can then carry that group between catalytic sites or transfer it to a substrate. This arrangement links the structure of the metabolite with the enzyme’s ability to coordinate sequential catalytic steps.
After bicarbonate activation and carboxyl-group attachment, the biotin-containing compound serves as an intermediary within the reaction sequence. Its attached carboxyl group can move between catalytic sites before reaching the final substrate. This carrier function helps explain how one biochemical reaction can coordinate chemically distinct steps within a carboxylation process.
These compounds are especially relevant to pathways involving fatty acid synthesis, amino acid metabolism, and energy production. Their participation connects biotin-dependent carboxylation with several major areas of biochemical metabolism. Examining them can therefore help researchers relate individual enzyme reactions to broader changes in cellular metabolic organization and regulation.
Analysis of these metabolites can clarify how catalytic sites coordinate bicarbonate activation, biotin carboxylation, and transfer of the carboxyl group to a substrate. It also helps researchers investigate metabolic regulation by connecting reaction intermediates with pathway activity. These insights are useful for interpreting how biochemical systems organize multistep enzymatic reactions.
Their importance follows from the dependence of several metabolic pathways on effective biotin utilization. When biotin use is impaired, the affected carboxylation reactions can have consequences for fatty acid synthesis, amino acid metabolism, and energy production. Studying these compounds therefore provides biochemical context for nutritional issues, inherited disorders, and related metabolic research.