Rxivist logo

Biotinylation by antibody recognition - A novel method for proximity labeling

By Daniel Z Bar, Kathleen Atkatsh, Urraca Tavarez, Michael R Erdos, Yosef Gruenbaum, Francis S Collins

Posted 11 Aug 2016
bioRxiv DOI: 10.1101/069187 (published DOI: 10.1038/nmeth.4533)

Identification of protein-protein interactions is a major goal of biological research. Despite technical advances over the last two decades, important but still largely unsolved challenges include the high-throughput detection of interactions directly from primary tissue and the identification of interactors of insoluble proteins that form higher-order structures. We have developed a novel, proximity-based labeling approach that uses antibodies to guide biotin deposition onto adjacent proteins in fixed cells and primary tissues. We used this method to profile the dynamic interactome of lamin A/C in multiple cell and tissue types under various treatment conditions. Our results suggest a considerable variation in the composition of the nuclear envelope of different tissues. Of note, DNA damage response proteins Ku70 and Ku80 are more abundant in the vicinity of lamin A/C after thermal stress. This increased affinity also applies to the progerin isoform, potentially contributing to the premature aging phenotype of Hutchinson-Gilford progeria syndrome. The ability to detect protein-protein interactions in intact tissues, and to compare affinities quantitatively under different conditions or in the presence of disease mutations, can provide a new window into cell biology and disease pathogenesis.

Download data

  • Downloaded 2,555 times
  • Download rankings, all-time:
    • Site-wide: 6,652
    • In biochemistry: 112
  • Year to date:
    • Site-wide: 47,815
  • Since beginning of last month:
    • Site-wide: 37,529

Altmetric data

Downloads over time

Distribution of downloads per paper, site-wide