Effects of temperature and monovalent cations on activity and quaternary structure of tryptophanase.
Effects of temperature and monovalent cations on the activity and the quaternary structure of tryptophanase of Escherichia coli were studied. The conversion of the apoenzyme into the active holoenzyme was attained at 30 degrees C in Tris-HCl buffer (pH 8.0) containing pyridoxal-P and K+, while no conversion occurred at 5 degrees C. The active holoenzyme thus formed was stable even at 5 degrees C, as long as the cation was present. When K+ was absent, however, the active enzyme gradually lost the activity upon chilling to 5 degrees C. The HPLC gel filtration analysis of the active holoenzyme and the low temperature-inactivated enzyme species revealed that the tetrameric holoenzyme dissociated into the dimeric apoenzyme concomitant with the low temperature-induced inactivation at 5 degrees C. The results of HPLC experiments together with other available evidence also suggest that the inactive tetrameric holoenzyme was first formed from the dimeric apoenzyme and pyridoxal-P prior to the formation of the active holoenzyme and that the cation promoted the conversion of the inactive holoenzyme into the active holoenzyme rather than being involved in the conversion of the apoenzyme and pyridoxal-P into the holoenzyme. Among various cations tested for the above effects, NH4+ exhibited the largest effect and K+ the second.
D000641:Ammonia / Q000494:pharmacology*; D002414:Cations, Monovalent; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000201:enzymology*; D008190:Lyases / Q000378:metabolism*; D046911:Macromolecular Substances; D011188:Potassium / Q000494:pharmacology*; D011732:Pyridoxal Phosphate / Q000378:metabolism; D012964:Sodium / Q000494:pharmacology*; D013696:Temperature; D014368:Tryptophanase / Q000378:metabolism*
D002414:Cations, Monovalent; D046911:Macromolecular Substances; D011732:Pyridoxal Phosphate; D000641:Ammonia; D012964:Sodium; D008190:Lyases; D014368:Tryptophanase; D011188:Potassium