《Thiol-Cleavable Biotin for Chemical and Enzymatic Biotinylation and Its Application to Mitochondrial TurboID Proteomics》 was written by Li, Haorong; Frankenfield, Ashley M.; Houston, Ryan; Sekine, Shiori; Hao, Ling. COA of Formula: C10H16N2O3SThis research focused onTurboID; cleavable biotin; mitochondrion; proximity labeling; streptavidin. The article conveys some information:
Protein biotinylation via chem. or enzymic reactions is often coupled with streptavidin-based enrichment and on-bead digestion in numerous biol. applications. However, the popular on-bead digestion method faces major challenges of streptavidin contamination, overwhelming signals from endogenous biotinylated proteins, the lost information on biotinylation sites, and limited sequence coverage of enriched proteins. Here, we explored thiol-cleavable biotin as an alternative approach to elute biotinylated proteins from streptavidin-coated beads for both chem. biotinylation and biotin ligase-based proximity labeling. All possible amino acid sites for biotinylation were thoroughly evaluated in addition to the primary lysine residue. We found that biotinylation at lysine residues notably reduces the trypsin digestion efficiency, which can be mitigated by the thiol-cleavable biotinylation method. We then evaluated the applicability of thiol-cleavable biotin as a substrate for proximity labeling in living cells, where TurboID biotin ligase was engineered onto the mitochondrial inner membrane facing the mitochondrial matrix. As a proof-of-principle study, thiol-cleavable biotin-assisted TurboID proteomics achieved remarkable intraorganelle spatial resolution with significantly enriched proteins localized in the mitochondrial inner membrane and mitochondrial matrix. In the experiment, the researchers used many compounds, for example, 5-((3aS,4S,6aR)-2-Oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoic acid(cas: 58-85-5COA of Formula: C10H16N2O3S)
5-((3aS,4S,6aR)-2-Oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoic acid(cas: 58-85-5) may be used to elute proteins from avidin/streptavidin resins. It has been used for culturing of oligodendrocytes.COA of Formula: C10H16N2O3S And it has been used as a vitamin supplement for the growth of Bacillus species.
Referemce:
Imidazole – Wikipedia,
Imidazole | C3H4N2 – PubChem