The Serine/Threonine Protein Phosphatase 2A (PP2A) Regulates Syk Activity in Human Platelets
Abstract
:1. Introduction
2. Results
2.1. PP2A Inhibition, but Not PP1 Inhibition, Results in Stoichiometric Syk S297 Phosphorylation in Platelets
2.2. Syk S297 Phosphorylation Induced by PP2A Inhibition Is Only Minimally Impaired by Syk or PKC Inhibitors
2.3. PP2A Inhibition Significantly Reduces Cvx-Stimulated Syk Y525/526 Phosphorylation (Kinase Domain)/Activity but Does Not Inhibit Cvx-Stimulated Syk S297 Phosphorylation (Interdomain-B)
2.4. Extended PP2A Inhibition Prevents Down-Regulation of and Prolongs Cvx-Stimulated Syk S297 Phosphorylation
3. Discussion
4. Materials and Methods
4.1. Blood Donors and Ethics Approval
4.2. Preparation and Stimulation of Human Platelets
4.3. SDS-PAGE, Zn2+ Phos-tagtm -SDS-PAGE and Western Blot Analysis
4.4. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ADP | Adenosine diphosphate |
BLNK | B-cell linker protein |
BTK | Bruton tyrosine kinase |
cAMP | Cyclic adenosine monophosphate |
cGMP | Cyclic guanosine monophosphate |
Clec-2 | C-type lectin 2 |
Cvx | Convulxin |
GFX | GF109203X; 2-[1-(3-Dimethylaminopropyl)indol-3-yl]-3-(indol-3-yl) maleimide |
ITAM | Immunoreceptor tyrosine-based activation motif |
LAT | Linker adaptor for T-cells |
PDBu | Phorbol 12, 13-dibutyrate |
PKC | Protein kinase C |
PLCγ2 | Phospholipase Cγ2 |
PRT | PRT060318; 2-(1R,2S)-2-aminocyclohexylamino)-4-(m-tolylamino)pyrimidine-5-carboxamide |
S | Serine |
Syk | Spleen tyrosine kinase |
T | Threonine |
TxA2 | Thromboxane A2 |
vWF | Von Willebrand factor |
Y | Tyrosine |
C | Control |
PP2A | Protein phosphatase 2 A |
PP1 | Protein phosphatase 1 |
OA | Okadaic acid |
EtOH | Ethanol |
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Makhoul, S.; Kumm, E.; Zhang, P.; Walter, U.; Jurk, K. The Serine/Threonine Protein Phosphatase 2A (PP2A) Regulates Syk Activity in Human Platelets. Int. J. Mol. Sci. 2020, 21, 8939. https://doi.org/10.3390/ijms21238939
Makhoul S, Kumm E, Zhang P, Walter U, Jurk K. The Serine/Threonine Protein Phosphatase 2A (PP2A) Regulates Syk Activity in Human Platelets. International Journal of Molecular Sciences. 2020; 21(23):8939. https://doi.org/10.3390/ijms21238939
Chicago/Turabian StyleMakhoul, Stephanie, Elena Kumm, Pengyu Zhang, Ulrich Walter, and Kerstin Jurk. 2020. "The Serine/Threonine Protein Phosphatase 2A (PP2A) Regulates Syk Activity in Human Platelets" International Journal of Molecular Sciences 21, no. 23: 8939. https://doi.org/10.3390/ijms21238939