Intrinsic Fluorescence of the Active and the Inactive Functional Forms of Human Thymidylate Synthase.
fluorescence
intrinsic protein fluorescence
protein functional forms
proteins
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
14 05 2021
14 05 2021
Historique:
revised:
04
02
2021
received:
19
10
2020
pubmed:
9
2
2021
medline:
15
12
2021
entrez:
8
2
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 33554411
doi: 10.1002/cbic.202000722
doi:
Substances chimiques
Deoxyuracil Nucleotides
0
2'-deoxyuridylic acid
964-26-1
Thymidylate Synthase
EC 2.1.1.45
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1800-1810Subventions
Organisme : Italian Association for Cancer Research (AIRC)
ID : IG 10474
Organisme : Italian Association for Cancer Research (AIRC)
ID : IG 16977
Informations de copyright
© 2021 Wiley-VCH GmbH.
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