The effect of divalent cations on the interaction between calf spleen profilin and different actins.
The interaction between calf spleen profilin and actin depends critically on the status of the C-terminus of the actin, and in the case of profilin, the C-terminus is of great importance for the physiochemical behaviour of the protein. Both proteins easily lose their C-terminal amino acids during the preparation, and special care has to be taken to ensure the isolation of the proteins in the intact form. Another factor that may seriously influence the study of the interaction of profilin with actin is the presence of varying amounts of an activity that causes an apparent stabilization of the complex even at later stages of its purification. We have found conditions for the isolation of intact profilin and actin, and studied the interaction between the two proteins, including the determination of the Kdiss for the complex formed under various ionic conditions. The complex formed between profilin and actin from calf spleen was found to be significantly stronger (Kdiss less than or equal to 10(-8) M in 50 mM KCl, and Kdiss = 4.10(-7) M in 50 mM KCl, 1 mM MgCl2) than that formed between profilin and muscle alpha-actin (Kdiss = 10(-6) M in 50 mM KCl, +/- 1 mM MgCl2). The profilactin complex formed in the mammalian system was stronger than the complex formed between Acanthamoeba actin and the profilin-like protein isolated from this organism. Analysis of the formation of the calf spleen complex in the presence of varying concentrations of divalent cations gave evidence for the presence of a high-affinity divalent-cation-binding site on the spleen actin (beta, gamma) which appears to regulate the interaction with profilin.
Larsson
H
H
Department of Zoological Cell Biology, Wenner-Gren Institute, University of Stockholm, Sweden.
Lindberg
U
U
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology*; D002413:Cations, Divalent; D002417:Cattle; D003285:Contractile Proteins / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology*; D015636:Magnesium Chloride; D008840:Microfilament Proteins / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism; D051304:Profilins; D011506:Proteins / Q000302:isolation & purification; D011817:Rabbits; D013154:Spleen / Q000378:metabolism*; D014783:Viscosity
D000199:Actins; D002413:Cations, Divalent; D003285:Contractile Proteins; D008840:Microfilament Proteins; D051304:Profilins; D011506:Proteins; C024782:profilactin; D015636:Magnesium Chloride; D008274:Magnesium; D002122:Calcium Chloride