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Electrophoretic demonstration of interactions between group specific component (vitamin D binding protein), actin, and 25‐hydroxycholecalciferol
Author(s) -
Emerson David L.,
Galbraith Robert M.,
Arnaud Philippe
Publication year - 1984
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150050104
Subject(s) - vitamin d binding protein , actin , vitamin d and neurology , isoelectric focusing , chemistry , isoelectric point , electrophoresis , immunofixation , vitamin , biochemistry , biology , genetics , monoclonal , antibody , monoclonal antibody , enzyme , endocrinology
Group specific component (Gc) is known to interact with both vitamin D 3 metabolites and actin. The present study was undertaken to determine if analytical isoelectric focusing could form the basis of a simple and reliable method for discriminating native Gc and complexes with 25‐hydroxycholecalciferol (25(OH)D 3 ) from those formed with actin and with both ligands. Increasing amounts of G‐actin and/or 25(OH)D 3 were added to purified Gc of both Gc1 and Gc2 phenotypes. Actin and 25(OH)D 3 interacted independently and simultaneously with native Gc, giving rise to three different complexes of increasing acidity: Gc‐25(OH)D 3 , Gc‐actin, and actin‐Gc‐25(OH)D 3 , which were clearly resolved from the native Gc protein and from each other. In addition, the inherent differences in microheterogeneity and polymorphism between the two major phenotypes examined (Gc1 and Gc2) were also retained in their respective complexes. Similar results were obtained upon addition of 25(OH)D 3 or actin to whole human serum, the bands corresponding to native or complexed Gc being recognized in this case by print immunofixation. These results indicate that complexes formed between Gc protein and both 25(OH)D 3 and actin can be clearly detected and resolved by electrophoretic procedures.