Multidomain proteins: conformational dynamics and foldingCopyright: Fitter
Large scale motions of globular domains are often crucial for the activity of multidomain proteins. We have employed single-molecule Förster resonance energy transfer (smFRET) for the characterization of large scale domain motions in the case of Phosphoglycerate kinase (PGK). PGK is a key enzyme in glycolysis which has a widely open domain structure with a hinge near the active centre between the two domains.
In prokaryotic and even more in eukaryotic cells the predominant fraction of the whole proteome belongs to the class of multidomain proteins. However, because of methodical reasons our existing knowledge about mechanisms and principles of protein folding results mainly from studies on smaller singledomain proteins. Therefore we investigated folding of PGK by single molecule FRET with dyes attached in the N- and C-domains. In addition to inter-domain distances we currently map several intra-domain distances for different folding states. This will provide us with a more complete picture about the sequential order of folding/unfolding transitions of individual domains.
Michele Cerminara, Antonie Schöne, Ilona Ritter, Matteo Gabba and Jörg Fitter
Mapping Multiple Distances in a Multidomain Protein for the Identification of Folding Intermediates
Biophys. J, 118, 1–10 (2020)
D. Kempe, M. Cerminara, S. Poblete, A. Schöne, M. Gabba, and J. Fitter
Single-molecule FRET measurements in additive-enriched aqueous solutions
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M. Gabba, S. Poblete, T. Rosenkranz, A. Katranidis, D. Kempe, T. Züchner, R.G. Winkler, G. Gompper, and J. Fitter
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Biophys. J., 107, 1913-1923, (2014)
T. Rosenkranz, R. Schlesinger, M. Gabba, J. Fitter
Native and unfolded states of a phosphoglycerate kinase studied by single molecule FRET
ChemPhysChem, 12, 704-710, (2011)