Kalbitzer, Hans Robert and Spoerner, Michael and Ganser, Petra and Hozsa, Constantin and Kremer, Werner (2009) Fundamental Link between Folding States and Functional States of Proteins. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 131 (46). pp. 16714-16719. ISSN 0002-7863,
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Folding and function of proteins are two aspects of proteins which are usually considered as basically unrelated phenomena that are optimized by evolution independently. From the funnel model of folding/unfolding and the associated energy landscape, we infer the paradigm that the minimum number of folding intermediates is determined by the number of all functional states of a protein ("essential" folding intermediates). Here, we demonstrate the supposed fundamental link using the Ras protein complexed with the GTP analogue GppNHp that occurs in two structural states coexisting in solution. State 2 was shown earlier to represent the effector interacting state, and the function of state 1 was hitherto unknown. By (31)P NMR spectroscopy, we demonstrate that state 1 represents the conformation interacting with guanine nucleotide exchange factors (GEFs). Denaturation experiments of the protein with a chaotropic reagent show that both functional states coexist during folding and unfolding. Application of high pressure represents another perturbation of the energy landscape, leading to an increased population of the state 1 as observed by NMR spectroscopy. The specific volume difference between the two states Delta V(12) is 17.2 +/- 0.5 mL mol(-1), indicating that state 1 represents a more open conformation of the protein. The free energies of stabilization for state 1 and state 2 at 278 K can be determined as 8.3 and 9.8 kJ mol(-1), respectively.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | SMALL MOLECULES; RAS; BINDING; PRESSURE; DYNAMICS; AGGREGATION; GTP; POLYPEPTIDE; TRANSITIONS; EXPRESSION; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Dr. Hans Robert Kalbitzer Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Remco Sprangers |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 01 Sep 2020 11:09 |
| Last Modified: | 01 Sep 2020 11:09 |
| URI: | https://pred.uni-regensburg.de/id/eprint/28108 |
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