Single step purification of rat liver aldolase using immobilized artificial membrane chromatography.
Aldolase B is a peripheral membrane protein. Immobilized artificial membrane (IAM) surfaces were used to purify rat liver aldolase B in a single chromatographic step. Selective elution required dipalmitoylphosphatidylcholine (DPPC) to be included in the mobile phase. Selective elution of aldolase from the IAM column when DPPC (0.2 mM) was added to the mobile phase indicates that DPPC was an affinity displacing ligand for this membrane associated protein. Since tissue preparation involved only homogenization and centrifugation, the single step purification of aldolase B using IAM chromatography is a very convenient method. The IAM stationary phase (1.5 g) has a loading capacity of at least 4.39 mg total protein from rat liver homogenates and typically approximately 17.7 microg of pure aldolase in a single step from approximately 60 mg wet weight rat liver cytosol can be obtained.
Liu
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Department of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy, Purdue University, West Lafayette, IN 47907, USA.
Cohen
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Pidgeon
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D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D051381:Rats; D017386:Sequence Homology, Amino Acid
D008567:Membranes, Artificial; D005634:Fructose-Bisphosphate Aldolase