Separation of fodrin subunits by affinity chromatography on calmodulin-Sepharose.
Spectrin is composed of two nonidentical subunits, with the 240-kDa subunit of nonerythroid spectrin (fodrin) able to bind calmodulin (CaM) Ca2+-dependently. It was found that in the presence of chaotropic salts this binding site was still expressed, although the subunits of fodrin were dissociated. This has been exploited for separating the fodrin subunits rapidly and quantitatively by affinity chromatography on calmodulin-Sepharose. When bovine fodrin was dissolved in 2 M KI + 1 mM Ca2+ and applied to CaM-Sepharose the beta subunit (235-kDa) passed through unretarded whereas the alpha subunit (240-kDa) bound and could be eluted with ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid. These subunits would reform the intact molecule when mixed and dialyzed.
D000818:Animals; D001923:Brain Chemistry; D002147:Calmodulin; D002352:Carrier Proteins / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity; D006863:Hydrogen-Ion Concentration; D008840:Microfilament Proteins / Q000302:isolation & purification*; D008854:Microscopy, Electron; D011485:Protein Binding; D012685:Sepharose; D013049:Spectrin
D002147:Calmodulin; D002352:Carrier Proteins; D008840:Microfilament Proteins; D012685:Sepharose; D013049:Spectrin