• T cell receptor-dependent S-acylation of ZAP-70 controls activation of T cells.

T cell receptor-dependent S-acylation of ZAP-70 controls activation of T cells.

The Journal of biological chemistry (2021-01-23)
Ritika Tewari, Bieerkehazhi Shayahati, Ying Fan, Askar M Akimzhanov

ZAP-70 is a tyrosine kinase essential for T cell immune responses. Upon engagement of the T cell receptor (TCR), ZAP-70 is recruited to the specialized plasma membrane domains, becomes activated, and is released to phosphorylate its laterally segregated targets. A shift in ZAP-70 distribution at the plasma membrane is recognized as a critical step in TCR signal transduction and amplification. However, the molecular mechanism supporting stimulation-dependent plasma membrane compartmentalization of ZAP-70 remains poorly understood. In this study, we identified previously uncharacterized lipidation (S-acylation) of ZAP-70 using Acyl-Biotin Exchange assay, a technique that selectively captures S-acylated proteins. We found that this posttranslational modification of ZAP-70 is dispensable for its enzymatic activity. However, the lipidation-deficient mutant of ZAP-70 failed to propagate the TCR pathway suggesting that S-acylation is essential for ZAP-70 interaction with its protein substrates. The kinetics of ZAP-70 S-acylation were consistent with TCR signaling events indicating that agonist-induced S-acylation is a part of the signaling mechanism controlling T cell activation and function. Taken together, our results suggest that TCR-induced S-acylation of ZAP-70 can serve as a critical regulator of T cell-mediated immunity.

Product Number
Product Description

Anti-Lck Antibody, clone 3A5, clone 3A5, Upstate®, from mouse
Poly-L-lysine solution, 0.01%, sterile-filtered, BioReagent, suitable for cell culture
Phosphatase Inhibitor Cocktail 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)
Hydroxylamine hydrochloride, ReagentPlus®, 99%
cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial
Anti-ZAP-70 Antibody, clone 2F3.2, clone 2F3.2, Upstate®, from mouse
n-Dodecyl β-D-maltoside, ≥98% (GC)
S-Methyl methanethiosulfonate, 97%