RT Journal Article
SR Electronic
T1 Diffusional and chemical control in the tyrosine kinase network of platelet CLEC-2 signalling
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 529859
DO 10.1101/529859
A1 Alexey Martyanov
A1 Fyodor A Balabin
A1 Joanne L Dunster
A1 Mikhail Aleksandrovich Panteleev
A1 Jonathan Gibbins
A1 Anastasia N Sveshnikova
YR 2019
UL http://biorxiv.org/content/early/2019/01/24/529859.abstract
AB C-type lectin-like receptor 2 (CLEC-2) is platelet membrane glycoprotein implicated in maintenance of blood vessel integrity and development of lymphatics. Organization and regulation of tyrosine kinase signalling network associated with CLEC-2 is poorly understood. Here we aimed to investigate CLEC-2 signal transduction using computational systems biology methods in combination with experimental approaches. We developed a 3D/stochastic multicompartmental computational model of CLEC-2 signalling, which was built around membrane-associated signalosome formation. The model predicted that both Syk and Src-family kinases phosphorylate CLEC-2, and that CLEC-2 ligation induces cytosolic calcium spiking. This was experimentally confirmed using flow cytometry and TIRF microscopy, respectively. Sensitivity analysis suggested that the CLEC-2 translocation to the signalosome region is one of the rate-limiting steps in the signal transduction process. In agreement with this prediction, CLEC-2-induced platelet activation was strongly temperature-dependent (unlike that mediated by G-protein coupled receptors) and was delayed by lipid raft disruption. Our results suggest a revised picture of the CLEC-2 signal transduction network functioning that emphasizes the crucial role of lipid raft structural rearrangement followed by tyrosine kinase feedback interplay.