PT - JOURNAL ARTICLE AU - Daniel Z Bar AU - Kathleen Atkatsh AU - Urraca Tavarez AU - Michael R Erdos AU - Yosef Gruenbaum AU - Francis S Collins TI - Biotinylation by antibody recognition - A novel method for proximity labeling AID - 10.1101/069187 DP - 2016 Jan 01 TA - bioRxiv PG - 069187 4099 - http://biorxiv.org/content/early/2016/08/11/069187.short 4100 - http://biorxiv.org/content/early/2016/08/11/069187.full AB - 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.