Chromatofocusing in the purification and separation of apo- and holo-(vitamin D-binding protein)
- 1 August 1985
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 229 (3) , 669-674
- https://doi.org/10.1042/bj2290669
Abstract
Chromatofocusing was used to purify the vitamin D-binding protein (DBP) from pig plasma in a procedure that consisted of an initial DEAE-cellulose chromatography followed by DEAE-Sephadex chromatography, with final purification by chromatofocusing. The protein was purified 184-fold over its concentration in plasma. When the plasma was labelled with a tracer concentration of [3H]calcidiol, it was apparent that holo- and apo-DBP did not co-chromatograph on chromatofocusing. The separation of these two forms of DBP on chromatofocusing was verified by using purified apo-DBP mixed with either a tracer or a saturating concentration of calcidiol. This separation was consistent with differences observed in their isoelectric points. The ability to separate apo and holo forms of DBP should permit the study of their specific interactions with other binding proteins and help determine the physiological relevance of these interactions.This publication has 17 references indexed in Scilit:
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