Herbst, Ruth and Schäfer, Ute and Seckler, Robert (1997) Equilibrium intermediates in the reversible unfolding of firefly (Photinus pyralis) luciferase. JOURNAL OF BIOLOGICAL CHEMISTRY, 272 (11). pp. 7099-7105. ISSN 0021-9258, 1083-351X
Full text not available from this repository.Abstract
Firefly luciferase has been used as a model protein to study cotranslational and chaperone-assisted protein folding, We found conditions for reversible unfolding of luciferase in the absence of cellular factors, and we characterized denaturant-induced equilibrium unfolding transitions and refolding kinetics of the enzyme. Luciferase unfolding induced by guanidinium chloride at 10 degrees C can be described as a four-state equilibrium with two inactive intermediates highly populated around 1 and 3 M denaturant, The transitions occur around 0.3, 1.7, and 3.8 M denaturant, The fi ee energy of denaturation to the first inactive intermediate (Delta G(N=11)(0) = 15 +/- 3 kJ . mol(-1)) is small for a protein of 60 kDa, Fluorescence and circular dichroism spectra of the intermediates indicate that I-1 has a compact conformation, whereas aromatic side chains are highly exposed in the second intermediate, I-2, despite its high content of secondary structure. In the presence of a hydrophilic detergent, significant reactivation of luciferase is observed up to temperatures at which the native protein is unstable, Reactivation kinetics of luciferase are exceedingly slow and probably not limited by proline isomerization, as suggested by their independence from the time spent in the unfolded state.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ESCHERICHIA-COLI; RETICULOCYTE LYSATE; LUCIOLA-MINGRELICA; PROTEIN; AGGREGATION; MECHANISM; CLONING; SYSTEM; CHAINS; CDNA |
| Subjects: | 500 Science > 540 Chemistry & allied sciences 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 11 May 2023 09:49 |
| Last Modified: | 11 May 2023 09:49 |
| URI: | https://pred.uni-regensburg.de/id/eprint/50979 |
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