Characterization of the interaction between calpactin I and fodrin (non-erythroid spectrin).
Calpactin I, a calcium-binding protein associated with the membrane cytoskeleton, has been reported to bind to a calcium-dependent manner to fodrin, to certain phospholipids, and to F-actin. We have investigated the interaction between calpactin I and fodrin. Using a gel filtration assay, we observed one or more calpactin I molecules were bound calcium-dependently only at high concentrations of calpactin (greater than 1 microM), indicating that the interaction is of only moderate affinity. At higher concentrations of calpactin I, the calpactin coprecipitated with fodrin in a calcium-dependent manner. The molar ratio of calpactin to fodrin tetramer in the precipitate was greater than 25:1, indicating that the calpactin binds to a large number of sites. Moreover, the monomeric form of calpactin I (p36), which did not induce precipitation of fodrin, showed no evidence of saturation in its binding to fodrin even when more than 30 mol of p36 were bound per mole of fodrin tetramer. Several proteins other than fodrin, including clathrin, alpha-actinin, and neurofilament-H, also interacted calcium-dependently with calpactin I in the gel filtration assay. These results demonstrate that the interaction between calpactin and fodrin is not of high affinity, is not readily saturated, and is not specific for fodrin. Our results suggest that calpactin's interaction with fodrin is a particular example of a calcium-dependent, but promiscuous, binding of calpactin to proteins.
Cheney
R E
RE
Department of Anatomy and Neurobiology, Washington University Medical School, St. Louis, Missouri 63110.
Willard
M B
MB
D000818:Animals; D017302:Annexins; D001921:Brain / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002850:Chromatography, Gel; D004533:Egtazic Acid / Q000494:pharmacology; D007700:Kinetics; D008168:Lung / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008840:Microfilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D013049:Spectrin / Q000378:metabolism
D017302:Annexins; D002118:Calcium; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D004533:Egtazic Acid; D008565:Membrane Proteins; D008840:Microfilament Proteins; D013049:Spectrin