Purification of tumor mitochondrial malic enzyme by specific ligand affinity chromatography.
A two-step chromatographic procedure, based on a specific ligand-binding approach, for the purification of tumor NAD(P)(+)-dependent malic enzyme is described. The enzyme was purified to near homogeneity by extraction from mitochondria, negative cellulose phosphate chromatography, ammonium sulfate precipitation, and application of specific elution from a malate-agarose column. The rationale for the use of the affinity column is also described.
Teller
J K
JK
Department of Pharmacology, University of Wisconsin Medical School, Madison 53706.
Fahien
L A
LA
D000645:Ammonium Sulfate; D000818:Animals; D002482:Cellulose / Q000031:analogs & derivatives; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D008291:Malate Dehydrogenase / Q000302:isolation & purification*; D008293:Malates; D008297:Male; D008928:Mitochondria / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D012685:Sepharose
D008293:Malates; C030298:malic acid; D002482:Cellulose; D012685:Sepharose; C008881:phosphocellulose; D008291:Malate Dehydrogenase; D000645:Ammonium Sulfate