Zaiss, Katrin and Jaenicke, Rainer (1999) Thermodynamic study of phosphoglycerate kinase from Thermotoga maritima and its isolated domains: Reversible thermal unfolding monitored by differential scanning calorimetry and circular dichroism spectroscopy. BIOCHEMISTRY, 38 (14). pp. 4633-4639. ISSN 0006-2960,
Full text not available from this repository. (Request a copy)Abstract
The folding of phosphoglycerate kinase (PGK) from the hyperthermophilic bacterium Thermotoga maritima and its isolated N- and C-terminal domains (N1/2 and C1/2) was characterized by differential scanning calorimetry (DSC) and circular dichroism (CD) spectroscopy. At pH 3.0-4.0, reversible thermal denaturation of TmPGK occurred below 90 degrees C. The corresponding peaks in the partial molar heat capacity function were fitted by a four-state model, describing three well-defined unfolding transitions. Using CD spectroscopy, these are ascribed to the disruption of the domain interactions and subsequent sequential unfolding of the two domains. The isolated N-terminal domain unfolds reversibly between pH 3.0 and pH 4.0 to >90% and at pH 7.0 to about 70%. In contrast, the isolated engineered C-terminal domain only shows reversible thermal denaturation between pH 3.0 and pH 3.5. Neither N1/2 nor C1/2 obeys the simple two-state mechanism of unfolding. Instead, both unfold via a partially structured intermediate. In the case of N1/2, the intermediate exhibits native secondary structure and perturbed tertiary structure, whereas for C1/2 the intermediate could not be defined with certainty.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PROTEIN; STABILITY; DENATURATION; MACROMOLECULES; MUTANTS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 08 Nov 2022 10:34 |
| Last Modified: | 08 Nov 2022 10:34 |
| URI: | https://pred.uni-regensburg.de/id/eprint/48335 |
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