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A quantitative procedure for recovery of nucleoside phosphate ligands and preparation of apoprotein from holoproteins using blue dextran-Sepharose chromatography

1976; Elsevier BV; Volume: 72; Issue: 1-2 Linguagem: Inglês

10.1016/0003-2697(76)90532-7

1096-0309

Sioe Thung Thompson, Robert Cass, Earle Stellwagen,

RNA modifications and cancer

A simple chromatographic procedure is described for removal of strongly bound ligands from holoproteins whose ligand sites are constructed by an NAD-like domain. Application of such a holoprotein solution to a blue dextran-Sepharose chromatographic column equilibrated with a low ionic strength solvent results in recovery of the ligand and retention of the apoprotein. The apoprotein is then recovered by either batch or gradient elution with solvents having a high ionic strength. The quantitative removal of NAD from glyceraldehyde-3-phosphate dehydrogenase and the preparation of its apoprotein are illustrated.