title: Correlation between inhibition of cytoskeleton proteolysis and anti-vesiculation effect of calpeptin during A23187-induced activation of human platelets: are vesicles shed by filopod fragmentation?
abstract: Platelets were incubated in the presence of calpeptin to inhibit calpain-mediated cytoskeleton proteolysis during further activation by Ca2+ ionophore A23187. The appearance of filamin and myosin subfragments (93 kDa and 135 kDa, respectively) was inhibited by low calpeptin doses (1 microgram/ml). Higher doses (10-20 micrograms/ml) were required to completely inhibit talin and filamin degradation. Vesiculation strongly depended on cytoskeleton proteolysis and was reduced by 60% when platelets were preincubated with 10 micrograms/ml calpeptin. Activated platelets bore longer and more filopods when pretreated with calpeptin. Filopods were straight and regular when high calpeptin doses were used, whereas they were shorter and broader with bloated surfaces when calpeptin was omitted. Some bloated areas were also found in straight filopods. These results suggest that the cytoskeleton proteolysis, and more specifically filamin proteolysis, induced bloating of filopod surfaces, thus facilitating fragmentation of filopod into vesicles.
authors.affiliation: URA 530 CNRS, Université Montpellier II, CP 107, France.
authors.forename: F
authors.initials: F
authors.lastname: Bassé
authors.forename: P
authors.initials: P
authors.lastname: Gaffet
authors.forename: A
authors.initials: A
authors.lastname: Bienvenüe
mesh_terms: D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D000001:Calcimycin / Q000494:pharmacology*; D002154:Calpain / Q000037:antagonists & inhibitors; D003285:Contractile Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004151:Dipeptides / Q000494:pharmacology*; D064448:Filamins; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism; D015539:Platelet Activation / Q000187:drug effects*
chemical_list: D003285:Contractile Proteins; D004151:Dipeptides; D064448:Filamins; D008840:Microfilament Proteins; C071482:calpeptin; D000001:Calcimycin; D002154:Calpain