SlyD Accelerates trans-to-cis Prolyl Isomerization in a Mechanosignaling Protein under Load
| Autor: | Lorenz Rognoni, Ulrich Merkel, Abhigyan Sengupta, Matthias Rief, Gabriel Žoldák |
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| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Chemistry 030303 biophysics Force spectroscopy Isomerase Filamin single-molecule force spectroscopy Surfaces Coatings and Films 03 medical and health sciences Enzyme Materials Chemistry Biophysics Native state Proline single-molecule optical trap experiments Physical and Theoretical Chemistry Cytoskeleton Isomerization 030304 developmental biology |
| Zdroj: | Journal of Physical Chemistry B |
| Popis: | Prolyl isomerization is recognized as one of the key regulatory mechanisms, which plays a crucial role in cell signaling, ion channel gating, phage virus infection, and molecular timing. This isomerization is usually slow but often accelerated by an enzyme, called peptidyl–prolyl isomerase (PPIase). In the current project, we investigate using single-molecule force spectroscopy (SMFS) the impact of a bacterial PPIase, SlyD, on the cis–trans isomerization of the proline 2225 (P2225) in an isolated 20th domain of a cytoskeletal mechanosensing protein filamin-A (FlnA20). To explore the FlnA20–PPIase interaction, we have used multiple SMFS modes, like constant velocity, constant distance, and jumping trap experiments. In our previous study, we reported the unique nature of the P2225, which is conserved in all naturally occurring filamins and can slowly (minutes) interconvert between cis–trans isomers, in absence of any PPIase. Our current results show a staggering 25-fold acceleration of the trans-to-cis isomerization rate in the presence of saturating SlyD concentration (7.25 μM) compared to the unenzymatic condition. A SlyD concentration-dependent depletion of the trans isomeric lifetime was also observed. Additionally, we observed that SlyD stabilizes the cis-isomer in the native state of FlnA20 by ∼2 kBT. This is the first single-molecule observation of the cis–trans isomerization catalysis by a PPIase in a mechanosensing protein. This work was supported by a Sonderforschungsbereich 863 A2 Grant and ZO 291/1-1 Grant from the Deutsche Forschungsgemeinschaft. M.R. acknowledges financial support through an instrument grant from the Institute for Advanced Study of Technische Universitat Munchen. |
| Databáze: | OpenAIRE |
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