Abstract
We have curated an in-depth subcellular proteomic map of primary human CD4+ T cells, divided into cytosolic, nuclear and membrane fractions generated by an optimized fractionation and HiRIEF-LC-MS/MS workflow for limited amounts of primary cells. The subcellular proteome of T cells was mapped under steady state conditions, as well as upon 15 min and 1 h of T cell receptor (TCR) stimulation, respectively. We quantified the subcellular distribution of 6,572 proteins and identified a subset of 237 potentially translocating proteins, including both well-known examples and novel ones. Microscopic validation confirmed the localization of selected proteins with previously known and unknown localization, respectively. We further provide the data in an easy-to-use web platform to facilitate re-use, as the data can be relevant for basic research as well as for clinical exploitation of T cells as therapeutic targets.
Original language | English (US) |
---|---|
Journal | Frontiers in Immunology |
Volume | 10 |
DOIs | |
State | Published - Nov 26 2019 |
Bibliographical note
KAUST Repository Item: Exported on 2020-10-01Acknowledgements: We would like to thank Rui Branca and Jessie Thorslund for assistance with the LC-MS. We acknowledge Henrik Johansson for fruitful discussions and support during MS sample preparation. We would also like to thank Jorrit Boekel for assistance with the Galaxy pipeline (all from department of Oncology and Pathology, Karolinska Institutet). Support by Fredrik Levander of NBIS (National Bioinformatics Infrastructure Sweden) is also acknowledged. We acknowledge the Cell Profiling Facility at Royal Institute of Technology, funded by Science for Life Laboratory and the National Microscopy Infrastructure, NMI (VR-RFI 2016-00968) for assistance in sample preparation and microscopy.