High affinity divalent cation binding to actin. Effect of low affinity salt binding.
Monomeric actin labeled with the fluorescent probe N-iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine (1,5-I-AEDANS-actin) displays a fast fluorescence intensity increase immediately upon addition of salt and then a slow fluorescence intensity change concurrent with Ca2+/Mg2+ exchange at the high affinity divalent cation binding site on actin. The fast change appears to reflect competitive binding of K+ at low affinity (nonspecific) sites and of Mg2+ or Ca2+ at low and intermediate affinity sites. Binding of cation at the low affinity sites (but apparently not at the intermediate affinity sites) results in an increase in k-Ca and k-Mg and thus a decrease in affinity for divalent cations at the high affinity site. The effect of Mg2+ on k-Ca is twice that of K+ for equal fractional saturations of the low affinity binding, and the effect of K+ and Mg2+ together on k-Ca reflects competitive binding at the low affinity sites. Thus the affinity and kinetics of divalent cation binding at the high affinity site of actin are significantly affected by concurrent cation binding at low affinity sites.
Selden
L A
LA
Research Service, Veterans Administration Medical Center, Albany, New York.
Estes
J E
JE
Gershman
L C
LC
D000199:Actins / Q000378:metabolism*; D000345:Affinity Labels; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002122:Calcium Chloride; D002413:Cations, Divalent / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D005456:Fluorescent Dyes; D008274:Magnesium; D015636:Magnesium Chloride; D009282:Naphthalenesulfonates; D011189:Potassium Chloride; D011817:Rabbits
D000199:Actins; D000345:Affinity Labels; D002413:Cations, Divalent; D002712:Chlorides; D005456:Fluorescent Dyes; D009282:Naphthalenesulfonates; D015636:Magnesium Chloride; D011189:Potassium Chloride; D008274:Magnesium; D002122:Calcium Chloride