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Resolution of transducin subunits by chromatography on blue sepharose

Item Type:Article
Title:Resolution of transducin subunits by chromatography on blue sepharose
Creators Name:Kleuss, C. and Pallast, M. and Brendel, S. and Rosenthal, W. and Schultz, G.
Abstract:The retinal guanine nucleotide-binding protein, transducin (TD), was subjected to chromatography on Blue Sepharose (BLS). A simple two-step protocol was developed, allowing the resolution of the alpha-subunit and the beta gamma-complex of the protein extracted from bovine retina by the use of a poorly hydrolysable GTP analogue. If TD was applied to BLS in a divalent cation-containing buffer, the beta gamma-complex did not bind to the resin, whereas the alpha-subunit was retained; elution of the latter was achieved by removing the divalent cation from the buffer. Binding of the alpha-subunit to BLS was not affected by nucleotides or by ADP ribosylation catalysed by bacterial toxins. However, adsorption of the alpha-subunit by BLS or by a strong cation exchanger (Mono S) depended strictly on divalent cations. In contrast to previous reports, the data suggest the formation of a complex between a sulphonyl residue of Cibacron Blue, a divalent metal ion, and the alpha-subunit as the relevant binding mechanism causing adsorption of the alpha-subunit to BLS.
Keywords:Ion Exchange Chromatography, Guanosine 5'-O-(3-Thiotriphosphate), Guanosine Triphosphate, Membrane Proteins, Retina, Sepharose, Sulfur Radioisotopes, Thionucleotides, Transducin, Animals, Cattle
Source:Journal of chromatography
Page Range:281-289
Date:16 October 1987
Official Publication:https://doi.org/10.1016/S0021-9673(01)92625-1
PubMed:View item in PubMed

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