Mechanisms of cold-induced platelet actin assembly.
Various agonists but also chilling cause blood platelets to increase cytosolic calcium, polymerize actin, and change shape. We report that cold increases barbed end nucleation sites in octyl glucoside-permeabilized platelets by 3-fold, enabling analysis of the intermediates of this response. Although chilling does not change polyphosphoinositide (ppI) levels, a ppI-binding peptide completely inhibits cold-induced nucleation. The C terminus of N-WASp, which inhibits the Arp2/3 complex, blocks nucleation by 40%; GDPbetaS, N17Rac and N17Cdc42 have no effects. Some gelsolin translocates to the detergent-insoluble cytoskeleton after cooling. Chilled platelets from gelsolin-deficient mice have approximately 50% fewer new actin nuclei compared with platelets from wild-type mice. EGTA completely inhibits gelsolin translocation into the cytoskeleton, and the small amount of gelsolin initially there becomes soluble. Chilling releases adducin from the detergent-resistant cytoskeleton. We conclude that platelet actin filament assembly induced by cooling involves ppI-mediated actin filament barbed end uncapping and de novo nucleation independently of surface receptors or downstream signaling intermediates besides calcium. The actin-related changes occur in platelets at temperatures below 37 degrees C, suggesting that the platelet may be more activable at temperatures at the body surface than at core temperature, thereby favoring superficial hemostasis over internal thrombosis.
Hoffmeister
K M
KM
Division of Hematology, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Ave., Boston, MA 02115, USA.
Falet
H
H
Toker
A
A
Barkalow
K L
KL
Stossel
T P
TP
Hartwig
J H
JH
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism / Q000648:ultrastructure*; D003080:Cold Temperature; D003598:Cytoskeletal Proteins; D020558:GTP Phosphohydrolases / Q000378:metabolism; D018260:Gelsolin / Q000378:metabolism; D005960:Glucosides; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D006487:Hemostasis; D006801:Humans; D051379:Mice; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D011108:Polymers; D011817:Rabbits; D013873:Thionucleotides / Q000378:metabolism; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal
C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; C495927:Actr2 protein, mouse; C495929:Actr3 protein, mouse; D003598:Cytoskeletal Proteins; D018260:Gelsolin; D005960:Glucosides; D009419:Nerve Tissue Proteins; D018129:Phosphatidylinositol Phosphates; D011108:Polymers; D013873:Thionucleotides; C495771:WASL protein, human; C495772:Wasl protein, mouse; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D006153:Guanosine Diphosphate; C018619:octyl-beta-D-glucoside; C023427:guanosine 5'-O-(2-thiodiphosphate); D020558:GTP Phosphohydrolases