Selective binding of gelsolin to actin monomers containing ADP.
The rate of reaction and the stoichiometry of binding between gelsolin and actin monomers depends on adenine nucleotides. In the presence of Ca2+ but not Mg2+, gelsolin retains the ability to sever actin filaments when incubated for more than 20 min with an excess of G-actin in the presence of ATP but loses severing activity within seconds when mixed with G-actin in ADP. Immunoprecipitation of gelsolin removes more actin from ADP than from ATP solutions. Monomeric ATP-actin in 2 mM MgCl2 and 150 mM KCl slowly destroys the filament-severing activity of gelsolin with kinetics that are first order in actin concentration and with an apparent bimolecular rate constant of 0.021 +/- 0.007 microM-1 s-1. Coincident with the slow complex formation in MgCl2, the actin bound to the calcium-sensitive actin binding domain of gelsolin hydrolyzes its ATP to ADP. These results suggest a further level of gelsolin regulation and a functional similarity between actin and GTP-binding proteins.
Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts.
L E
LE
Laham
J A
JA
Lamb
P G
PG
Allen
P A
PA
Janmey
D000199:Actins / Q000302:isolation & purification / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002850:Chromatography, Gel; D018260:Gelsolin; D006801:Humans; D007700:Kinetics; D008027:Light; D008274:Magnesium / Q000494:pharmacology; D008433:Mathematics; D008840:Microfilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008962:Models, Theoretical; D009132:Muscles / Q000378:metabolism; D011817:Rabbits; D012542:Scattering, Radiation
D000199:Actins; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D008274:Magnesium; D002118:Calcium