Proteomics

Dataset Information

0

Biotinylation by antibody recognition - A novel method for proximity labeling


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 showed our method to be specific and sensitive by labeling a mitochondrial matrix protein. Next, 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. The ability to detect proximal proteins and putative interactors 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.

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Fusion, LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Skeletal Muscle, Primary Cell, Adipose Tissue, Smooth Muscle, Hela Cell

SUBMITTER: Daniel Bar  

LAB HEAD: Francis Collins

PROVIDER: PXD004736 | Pride | 2017-10-17

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
3211.msf Msf
3211B1.msf Msf
3211B1.raw Raw
3211B2.msf Msf
3211B2.raw Raw
Items per page:
1 - 5 of 128
altmetric image

Publications

Biotinylation by antibody recognition-a method for proximity labeling.

Bar Daniel Z DZ   Atkatsh Kathleen K   Tavarez Urraca U   Erdos Michael R MR   Gruenbaum Yosef Y   Collins Francis S FS  

Nature methods 20171218 2


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  ...[more]

Similar Datasets

2018-11-04 | PXD009748 | Pride
2015-07-08 | E-GEOD-70618 | biostudies-arrayexpress
2017-09-29 | PXD006359 | Pride
2010-08-12 | E-GEOD-23609 | biostudies-arrayexpress
2022-02-22 | PXD028830 | Pride
2010-05-26 | E-GEOD-10452 | biostudies-arrayexpress
2010-05-06 | E-GEOD-14329 | biostudies-arrayexpress
2024-05-10 | PXD047121 | Pride
2014-12-15 | E-GEOD-53331 | biostudies-arrayexpress
2021-06-16 | PXD021622 | Pride