The use of 1-anilino-8-naphthalene sulfonate as fluorescent probe for conformational studies on ribulose-1,5-bisphosphate carboxylase.
The influence of Mg2+ ions and temperature on the structure of the enzyme ribulose-1,5-bisphosphate carboxylase was investigated using the fluorescent probe 1-anilino-8-naphthalene sulfonate (ANS). The binding of ANS to the enzyme molecule caused a significant increase of fluorescence emission which was further enhanced by the addition of Mg2+. The temperature dependence of the fluorescence emission indicated a conformational change of the enzyme between 12 and 24 degrees C. The Mg2+ and the temperature effects were additive. ANS itself did not change the conformation of the enzyme. The influence of the substrates carbon dioxide and ribulose-1,5-bisphosphate, and the effect of the pH of the medium and of a sulfhydryl reducing reagent on fluorescence emission were analysed.
D000817:Anilino Naphthalenesulfonates; D002262:Carboxy-Lyases; D004229:Dithiothreitol / Q000494:pharmacology; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008722:Methods; D011487:Protein Conformation / Q000187:drug effects; D012273:Ribulose-Bisphosphate Carboxylase; D012274:Ribulosephosphates / Q000494:pharmacology; D013696:Temperature
D000817:Anilino Naphthalenesulfonates; D012274:Ribulosephosphates; D002262:Carboxy-Lyases; D012273:Ribulose-Bisphosphate Carboxylase; D008274:Magnesium; D004229:Dithiothreitol