TY - JOUR
T1 - Quantitative analysis of protein-protein interactions and post-translational modifications in rare immune populations
AU - Avin, Ayelet
AU - Levy, Maayan
AU - Porat, Ziv
AU - Abramson, Jakub
N1 - We thank Dr. Eran Elinav for providing materials and technical support for the inflammasome experiments. The research in the Abramson lab is kindly supported by the by the European Research Council Consolidator Grant—ERC-2016-CoG-724821, Israel Science Foundation (722/14 and 1796/16); Sy Syms Foundation; Binational Science Foundation; Rising Tide Foundation; Maurice and Vivienne Wohl Charitable Foundation; Goodman Family Charitable Lead Annuity Trust; and Ruth and Samuel David Gameroff Family Foundation. J.A. is an incumbent of the Dr. Celia Zwillenberg-Fridman and Dr. Lutz Fridman Career Development Chair. Author contributions J.A. and A.A. designed the study and wrote the manuscript; A.A. performed most of the experimental work; A.A. and Z.P. preformed and designed the IFC analysis. M.L. helped with BMDM samples preparations. M.L., Z.P. and J.A. helped in performing, analyzing, and/or designing some of the experiments.
PY - 2017/12/1
Y1 - 2017/12/1
N2 - In spite of recent advances in proteomics, quantitative analyses of protein-protein interactions (PPIs) or post-translational modifications (PTMs) in rare cell populations remain challenging. This is in particular true for analyses of rare immune and/or stem cell populations that are directly isolated from humans or animal models, and which are often characterized by multiple surface markers. To overcome these limitations, here we have developed proximity ligation imaging cytometry (PLIC), a protocol for proteomic analysis of rare cells. Specifically, by employing PLIC on medullary thymic epithelial cells (mTECs), which serve as a paradigm for a rare immune population, we demonstrate that PLIC overcomes the inherent limitations of conventional proteomic approaches and enables a high-resolution detection and quantification of PPIs and PTMs at a single cell level.
AB - In spite of recent advances in proteomics, quantitative analyses of protein-protein interactions (PPIs) or post-translational modifications (PTMs) in rare cell populations remain challenging. This is in particular true for analyses of rare immune and/or stem cell populations that are directly isolated from humans or animal models, and which are often characterized by multiple surface markers. To overcome these limitations, here we have developed proximity ligation imaging cytometry (PLIC), a protocol for proteomic analysis of rare cells. Specifically, by employing PLIC on medullary thymic epithelial cells (mTECs), which serve as a paradigm for a rare immune population, we demonstrate that PLIC overcomes the inherent limitations of conventional proteomic approaches and enables a high-resolution detection and quantification of PPIs and PTMs at a single cell level.
UR - http://www.scopus.com/inward/record.url?scp=85034427570&partnerID=8YFLogxK
U2 - 10.1038/s41467-017-01808-6
DO - 10.1038/s41467-017-01808-6
M3 - مقالة
SN - 2041-1723
VL - 8
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 1524
ER -