Liu H, Cohen D E, Pidgeon C
Department of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy, Purdue University, West Lafayette, IN 47907, USA.
J Chromatogr B Biomed Sci Appl. 1997 Dec 5;703(1-2):53-62. doi: 10.1016/s0378-4347(97)00411-8.
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.
醛缩酶B是一种外周膜蛋白。固定化人工膜（IAM）表面用于在单一色谱步骤中纯化大鼠肝脏醛缩酶B。选择性洗脱需要在流动相中加入二棕榈酰磷脂酰胆碱（DPPC）。当向流动相中加入DPPC（0.2 mM）时，醛缩酶从IAM柱上的选择性洗脱表明DPPC是这种膜相关蛋白的亲和置换配体。由于组织制备仅涉及匀浆和离心，因此使用IAM色谱法对醛缩酶B进行单步纯化是一种非常方便的方法。IAM固定相（1.5 g）对大鼠肝脏匀浆中总蛋白的负载量至少为4.39 mg，通常从约60 mg湿重的大鼠肝脏胞质溶胶中一步可获得约17.7 μg的纯醛缩酶。
