Intrinsic Fluorescence of the Active and the Inactive Functional Forms of Human Thymidylate Synthase
| Autor: | Simone Vitiello, Monica Caselli, Glauco Ponterini, Maria Paola Costi, Matteo Santucci, Stefania Ferrari, Giorgia Pavesi |
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| Rok vydání: | 2021 |
| Předmět: |
Kinetics
Fluorescence Polarization Molecular Dynamics Simulation Intrinsic fluorescence 010402 general chemistry 01 natural sciences Biochemistry Thymidylate synthase chemistry.chemical_compound Humans Protein Structure Quaternary Molecular Biology chemistry.chemical_classification biology 010405 organic chemistry Cell growth Organic Chemistry A protein Thymidylate Synthase Fluorescence 0104 chemical sciences Monomer Enzyme chemistry Mutagenesis Site-Directed Biophysics biology.protein Molecular Medicine fluorescence · intrinsic protein fluorescence ·protein functional forms · proteins Deoxyuracil Nucleotides |
| Zdroj: | ChemBioChem. 22:1800-1810 |
| ISSN: | 1439-7633 1439-4227 |
| DOI: | 10.1002/cbic.202000722 |
| Popis: | The observables associated with protein intrinsic fluorescence - spectra, time decays, anisotropies - offer opportunities to monitor in real time and non-invasively a protein's functional form and its interchange with other forms with different functions. We employed these observables to sketch the fluorometric profiles of two functional forms of human thymidylate synthase (hTS), a homodimeric enzyme crucial for cell proliferation and thus targeted by anticancer drugs. The protein takes an active and an inactive form. Stabilization of the latter by peptides that, unlike classical hTS inhibitors, bind it at the monomer/monomer interface offers an alternative inhibition mechanism that promises to avoid the onset of drug resistance in anticancer therapy. The fluorescence features depicted herein can be used as tools to identify and quantify each of the two protein forms in solution, thus making it possible to investigate the kinetic and thermodynamic aspects of the active/inactive conformational interchange. Two examples of fluorometrically monitored interconversion kinetics are provided. |
| Databáze: | OpenAIRE |
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