Details

Autor(en) / Beteiligte

• Robinson, Jack B.
• Strottmann, James M.
• Wick, Donald G.
• Stellwagen, Earle

Titel

Affinity Chromatography in Nonionic Detergent Solutions

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 1980-10, Vol.77 (10), p.5847-5851

Ort / Verlag

United States: National Academy of Sciences of the United States of America

Erscheinungsjahr

1980

Quelle

EZB-FREE-00999 freely available EZB journals

Beschreibungen/Notizen

• Anionic dye affinity chromatography is commonly unproductive in the presence of nonionic detergents used to extract particulate proteins. Using lactate dehydrogenase as a model protein, Cibacron blue F3GA as a model dye, and Triton X-100 as a model detergent, we find that the dye is encapsulated in nonionic detergent micelles, rendering the dye incapable of ligation with the enzyme. However, the dye can be liberated from the micelles without altering the nonionic detergent concentration by addition of an anionic detergent, such as deoxycholate or sodium dodecyl sulfate, forming mixed anionic/nonionic micelles that displace the anionic dye. Encapsulation of the anionic detergents prevents their activity as protein denaturants. These observations have been successfully translated to the dye affinity chromatography of a detergent extract of brain particulate cyclic nucleotide phosphodiesterase.