A fluorescence study of conformational changes induced by substrate and temperature in bovine liver thiosulfate sulfurtransferase.
Structural transitions occurring in the range of 0-50 degrees C have been detected and studied in the enzyme thiosulfate sulfurtransferase (thiosulfate:cyanide sulfurtransferase, EC 2.8.1.1) by investigating both the intrinsic protein fluorescence and the fluorescence of covalently bound probes. The intrinsic fluorescence of the enzyme decreases sharply at 27 degrees C and the magnitude of this quenching is smaller for the sulfur-substituted enzyme (ES) than for the free enzyme (E), both of which are obligatory catalytic intermediates. The effect with ES is fully reversible and almost completely so with E. Fluorescence depolarization sudies with thiosulfate sulfurtransferase labeled at a number of different sites with the fluorosphore dimethylaminonaphthalene show a sharp increase in the polarization starting at 27 degrees C when the temperature/viscosity ratio is varied with temperature and no transition when the ratio is varied with glycerol. Enzyme activity show no transition at 20 degrees C but falls abruptly above 40 degrees C. Under the appropriate conditions, the 27 degrees C transition will lead to association of thiosulfate sulfurtransferase molecules and the appearance of turbidity. Both activity and fluorescence measurements support the idea that the ES form is significantly more stable than the E form. These results may result from changes in the interactions between the structural domains into which the single polypeptide chain of thiosulfate sulfurtransferase is folded.
D000818:Animals; D002417:Cattle; D003619:Dansyl Compounds; D004355:Drug Stability; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D008099:Liver / Q000201:enzymology*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013466:Sulfurtransferases / Q000378:metabolism*; D013696:Temperature; D013884:Thiosulfate Sulfurtransferase / Q000378:metabolism*
D003619:Dansyl Compounds; D005456:Fluorescent Dyes; D013466:Sulfurtransferases; D013884:Thiosulfate Sulfurtransferase; C029038:dansyl chloride