title: Soybean metal-binding proteins: calmodulin purification by hydrophobic interaction with polymer 3520.
abstract: Polymer 3520, a non-polar styrene divinylbenzene polymer, provides a simple way to purify calmodulin (CAM) from soybeans. This polymer, which selectively adsorbs CAM by hydrophobic interaction within the polymer matrix, contains no exchangeable groups; thus, interaction with CAM requires no Ca++ ions, and elution is achieved with 50% ethanol. Purification by this form of reversed-phase liquid chromatography is a substantial improvement over the conventional method, which requires high salt in elution buffers. CAM in soybean meal is first extracted with 80% ethanol in the presence of EGTA at room temperature and then chromatographed directly on a polymer 3520 column to yield pure CAM. Addition of non-ionic detergent (Nonidet P-40) to the ethanolic extract helps to separate extraneous proteins, lipids, sugars, and isoflavones. Such isolated CAM migrates as a single band during polyacrylamide gel electrophoresis and reversed-phase HPLC, and it retains activity stimulatory to phosphodiesterase.
authors.affiliation: Northern Regional Research Center, US Department of Agriculture, Peoria, IL 61604.
authors.forename: L C
authors.initials: LC
authors.lastname: Wang
authors.forename: R E
authors.initials: RE
authors.lastname: Peterson
authors.forename: K J
authors.initials: KJ
authors.lastname: Shang
authors.forename: A K
authors.initials: AK
authors.lastname: Ho
authors.forename: J A
authors.initials: JA
authors.lastname: Rothfus
mesh_terms: D000327:Adsorption; D002021:Buffers; D002147:Calmodulin / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol / Q000378:metabolism; D011137:Polystyrenes / Q000378:metabolism*; D011145:Polyvinyls / Q000378:metabolism*; D013025:Glycine max / Q000201:enzymology*
chemical_list: D002021:Buffers; D002147:Calmodulin; D011137:Polystyrenes; D011145:Polyvinyls; C051242:polymer 3520; D000431:Ethanol