Biotinylation by antibody recognition—a method for proximity labeling

• Published in: Nature methods Vol. 15; no. 2; pp. 127 - 133
• Main Authors: Bar, Daniel Z, Atkatsh, Kathleen, Tavarez, Urraca, Erdos, Michael R, Gruenbaum, Yosef, Collins, Francis S
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01-02-2018; Nature Publishing Group
• Subjects: 13; 13/1; 631/1647/2067; 631/1647/2230/2232; 631/1647/296; 631/45/475/2290; 82; 82/58; Animals; Antibodies - metabolism; Antigens; Bioinformatics; Biological Microscopy; Biological research; Biological Techniques; Biomedical Engineering/Biotechnology; Biotin; Biotin - metabolism; Biotinylation; Biotinylation - methods; Cellular proteins; Disease susceptibility; Gene mutation; Genetic aspects; HeLa Cells; Humans; Identification and classification; Innovations; Isotope Labeling - methods; Labeling; Life Sciences; Mass spectrometry; Mass spectroscopy; Mice; Mice, Inbred C57BL; Mitochondria; Mutation; Pathogenesis; Peptide mapping; Properties; Protein Interaction Mapping - methods; Proteins; Proteins - metabolism; Proteomics; Proteomics - methods; Proximity
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.4533

Proximity-based labeling represents a useful approach for mapping protein environment, but current methods for this are limited to application to cell lines. This approach is now extended to primary human tissues with a method that uses antibodies to guide proximity labeling.
The high-throughput detection of organelle composition and proteomic mapping of protein environment directly from primary tissue as well as the identification of interactors of insoluble proteins that form higher-order structures have remained challenges in biological research. We report a proximity-based labeling approach that uses an antibody to a target antigen to guide biotin deposition onto adjacent proteins in fixed cells and primary tissues, which allows proteins in close proximity to the target antigen to be captured and identified by mass spectrometry. We demonstrated the specificity and sensitivity of our method by examining the well-studied mitochondrial matrix. We then used the method to profile the dynamic interactome of lamin A/C in multiple cell and tissue types under various treatment conditions. The ability to detect proximal proteins and putative interactors in intact tissues, and to quantify changes caused by different conditions or in the presence of disease mutations, can provide a window into cell biology and disease pathogenesis.