Mass spectrometry locates local and allosteric conformational changes that occur on cofactor binding
Rebecca Beveridge1, Lukasz G Migas1, Karl A P Payne1
1Michael Barber Centre for Collaborative Mass Spectrometry, School of Chemistry, Centre for Synthetic Biology of Fine and Speciality Chemicals, Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK.
The Fdc1 enzyme
Area of Science:
- Biochemistry
- Enzymology
- Structural Biology
Background:
- Fdc1 is a decarboxylase enzyme requiring a unique prenylated FMN cofactor.
- Understanding cofactor binding's structural impact is crucial for enzyme function.
Purpose of the Study:
- To investigate the structural consequences of cofactor binding to the Fdc1 enzyme.
- To demonstrate the utility of native mass spectrometry techniques in studying protein-cofactor interactions.
Main Methods:
- Native top-down and bottom-up mass spectrometry (MS).
- Ion mobility mass spectrometry (IM-MS).
- Molecular dynamics (MD) simulations.
- Hydrogen-deuterium exchange mass spectrometry (HDX-MS).
Main Results:
- Cofactor binding stabilizes the Fdc1 enzyme structure.
- IM-MS revealed distinct conformational ensembles for apo and holo Fdc1.
- HDX-MS indicated widespread allosteric conformational changes upon cofactor binding.
Conclusions:
- Cofactor binding induces significant structural rearrangements in Fdc1.
- The study highlights the power of integrated MS techniques for characterizing protein dynamics and allostery.
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