quantitative kinetic study of the actin-bundling protein l-plastin and of its impact on actin turn-over定量的actin-bundling蛋白质l-plastin和动力学研究它对肌动蛋白人事变动的影响.pdfVIP

Quantitative Kinetic Study of the Actin-Bundling Protein L-Plastin and of Its Impact on Actin Turn-Over 1 1 1 ´ 2 ` Ziad Al Tanoury , Elisabeth Schaffner-Reckinger , Aliaksandr Halavatyi , Celine Hoffmann , Michele 1 1 1 1 1 Moes , Ermin Hadzic , Marie Catillon , Mikalai Yatskou , Evelyne Friederich * 1 Laboratory of Cytoskeleton and Cell Plasticity, Life Sciences Research Unit, University of Luxembourg, Luxembourg City, Luxembourg, 2 Laboratory of Plant Molecular ´ Biology, Public Research Center for Health (CRP-Sante), Strassen, Luxembourg Abstract Background: Initially detected in leukocytes and cancer cells derived from solid tissues, L-plastin/fimbrin belongs to a large family of actin crosslinkers and is considered as a marker for many cancers. Phosphorylation of L-plastin on residue Ser5 increases its F-actin binding activity and is required for L-plastin-mediated cell invasion. Methodology/Principal Findings: To study the kinetics of L-plastin and the impact of L-plastin Ser5 phosphorylation on L- plastin dynamics and actin turn-over in live cells, simian Vero cells were transfected with GFP-coupled WT-L-plastin, Ser5 substitution variants (S5/A, S5/E) or actin and analyzed by fluorescence recovery after photobleaching (FRAP). FRAP data were explored by mathematical modeling to estimate steady-state reaction parameters. We demonstrate that in Vero cell focal adhesions L-plastin undergoes rapid cycles of association/dissociation following a two-binding-state model. Phosphorylation of L-plastin increased its association r