A quantitative procedure for recovery of nucleoside phosphate ligands and preparation of apoprotein from holoproteins using blue dextran-Sepharose chromatography
✍ Scribed by Sioe Thung Thompson; Robert Cass; Earle Stellwagen
- Publisher
- Elsevier Science
- Year
- 1976
- Tongue
- English
- Weight
- 238 KB
- Volume
- 72
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A simple chromatographic procedure is described for removal of strongly bound ligands from holoproteins whose ligand sites are constructed by an NADlike 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-3phosphate dehydrogenase and the preparation of its apoprotein are illustrated.