A quantitative proteomic analysis of cofilin phosphorylation in myeloid cells and its modulation using the LIM kinase inhibitor Pyr1

Renaud Prudent (1) , Nathalie Demoncheaux (1) , Hélène Diemer (2) , Véronique Collin-Faure (3) , Reuben Kapur (4) , Fabrice Paublant (1) , Laurence Lafanechère (5) , Sarah Cianferani (2) , Thierry Rabilloud (3)

1 Cellipse SAS
2 LSMBO - Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg]
3 ProMD - Protéomique, Métaux et Différenciation
4 Departments of Medicine and Pediatrics [Indianapolis, IN, USA]
5 INSERM U823 - Institut d'oncologie/développement Albert Bonniot de Grenoble

LIM kinases are located at a strategic crossroad, downstream of several signaling pathways and upstream of effectors such as microtubules and the actin cytoskeleton. Cofilin is the only LIM kinases substrate that is well described to date, and its phosphorylation on serine 3 by LIM kinases controls cofilin actin-severing activity. Consequently, LIM kinases inhibition leads to actin cytoskeleton disorganization and blockade of cell motility, which makes this strategy attractive in anticancer treatments. LIMK has also been reported to be involved in pathways that are deregulated in hematologic malignancies, with little information regarding cofilin phosphorylation status. We have used proteomic approaches to investigate quantitatively and in detail the phosphorylation status of cofilin in myeloid tumor cell lines of murine and human origin. Our results show that under standard conditions, only a small fraction (10 to 30% depending on the cell line) of cofilin is phosphorylated (including serine 3 phosphorylation). In addition, after a pharmacological inhibition of LIM kinases, a residual cofilin phosphorylation is observed on serine 3. Interestingly, this 2D gel based proteomic study identified new phosphorylation sites on cofilin, such as threonine 63, tyrosine 82 and serine 108.

Domaines: Génomique, Transcriptomique et Protéomique [q-bio.GN]

Renaud Prudent, Nathalie Demoncheaux, Hélène Diemer, Véronique Collin-Faure, Reuben Kapur, et al.. A quantitative proteomic analysis of cofilin phosphorylation in myeloid cells and its modulation using the LIM kinase inhibitor Pyr1. PLoS ONE, 2018, 13 (12), pp.e0208979. ⟨10.1371/journal.pone.0208979⟩. ⟨hal-01961564⟩