Study on Escherichia coli alkaline phosphatase conformation by phosphorimetry in the presence of denaturant.
The influence of different denaturants on the phosphorescence spectrum and lifetime decay of Escherichia coli alkaline phosphatase (AP) was investigated. Phosphorescence intensity and lifetime of tryptophan residue (Trp-109) decrease upon addition of guanidine hydrochloride, ethylene diamine tetraacetic acid, and urea or decreasing acidity. The experiments show that AP undergoes different pathways with different denaturants and that the activation energy data, DeltaS degrees (not equal) and deltaH degrees (not equal) further confirm that there is a stable intermediate state between the folded and unfolded AP states in solution.
College of Food and Bioengineering, South China University of Technology, Guangzhou 510640, China.
Hai-Rong
HR
Zhang
Si-Yuan
SY
Guo
Lin
L
Li
Miao-Yan
MY
Cai
D000469:Alkaline Phosphatase / Q000737:chemistry*; D002151:Calorimetry; D004926:Escherichia coli / Q000201:enzymology*; D019791:Guanidine / Q000494:pharmacology*; D049449:Luminescence; D011487:Protein Conformation; D011489:Protein Denaturation; D013816:Thermodynamics; D014364:Tryptophan / Q000032:analysis
D014364:Tryptophan; D000469:Alkaline Phosphatase; D019791:Guanidine