Proximity Label Detection of Biotinylated Proteins

Studying protein subcellular distribution, assembly and interactions is critical to our understanding of fundamental biological processes. Proximity-dependent biotinylation (PDB) is a means for studying living cells and protein interactions. Protein-protein interaction (PPI) networks underlie all signaling pathways in cells and are typically detected by purifying and analyzing protein complexes from cell lysates. Proximity labeling (PL) is a technique for labeling endogenous interactions of specific proteins by gene fusion into promiscuous enzymes that catalyze the generation of diffusion-reactive species in living cells. Proximate biotinylation of proteins is an effective method for detecting protein-protein interactions. Most proximity labeling techniques use promiscuous biotin ligases or peroxidases fused to the protein of interest, enabling covalent biotin labeling of proteins. Interacting and neighboring proteins can then be captured and identified.

The Services

Alfa Chemistry's research team focuses on the innovation of adjacent biotinylated protein technology, which can be used for efficient protein-protein interaction detection. We provides efficient proximity biotinylation labeling services, including but not limited to:

• Proximity to biotinylated labels to detect protein interactions
• Design and improvement of detection scheme
• Analysis of biotin content in samples
• Sample handling and analysis
• Improvement of PDB enzyme properties
• Professional data processing
• Development of patterns for adjacent biotinylated protein labeling

Biotin ligase

Biotin is required by all living organisms and is produced by plants, fungi and most prokaryotes. Mammals do not produce biotin, and the uptake of biotin is mainly mediated by the multivitamin sodium transporter. Biotin acts as a covalently bound cofactor for biotin-dependent carboxylases in amino acid, fatty acid, carbohydrate, and energy metabolism. Biotin protein ligases (BPLs) are responsible for the covalent attachment of biotin to carboxylases and are present in all biological species. Biotin ligase has high specificity to carboxylase and plays an important role in the field of biotechnology.

Protein-protein interaction

Protein-protein interaction (PPI) refers to the process of combining two or more proteins, usually involved in important physiological processes. Proximity labeling is an efficient method for detecting protein-protein interactions. The biotinidase protein proximity labeling technology uses biotin ligase to biotinylate adjacent proteins. Separation by affinity between biotin-avidin/streptavidin combined with high-efficiency mass spectrometry analysis allows efficient identification of biotinylated proteins. Proximity labeling of proteins can be used to detect protein interactions, and it also provides new options for the isolation and purification of proteins on other cellular structures.

Versatile and efficient detection platform

• For efficient proximity biotinylation detection
• Biotinylation site analysis and identification
• Professional data analysis

Alfa Chemistry supports efficient proximity biotinylation detection services, which can detect efficient protein-protein interactions, accurately locate biotinylated proteins, and speed up the analysis of key proteins. If you are interested in our services, please contact us immediately.

References:

1. Marta. F.S.; et al. Protein−protein interaction detection in vitro and in cells by proximity biotinylation. J. Am. Chem. Soc. 2008, 130(29): 9251-9253.
2. Deepanksha. A.; et al. Establishment of proximity-dependent biotinylation approaches in different plant model systems. The Plant Cell. 2020, 32(11): 3388-3407.

※ It should be noted that our service is only used for research, not for clinical use.