@article {Bar069187, author = {Daniel Z Bar and Kathleen Atkatsh and Urraca Tavarez and Michael R Erdos and Yosef Gruenbaum and Francis S Collins}, title = {Biotinylation by antibody recognition - A novel method for proximity labeling}, elocation-id = {069187}, year = {2016}, doi = {10.1101/069187}, publisher = {Cold Spring Harbor Laboratory}, abstract = {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.}, URL = {https://www.biorxiv.org/content/early/2016/08/11/069187}, eprint = {https://www.biorxiv.org/content/early/2016/08/11/069187.full.pdf}, journal = {bioRxiv} }