Experiments suggest that simulations may overestimate electrostatic contributions to the mechanical stability of a fibronectin type III domain.
Steered molecular dynamics simulations have previously been used to investigate the mechanical properties of the extracellular matrix protein fibronectin. The simulations suggest that the mechanical stability of the tenth type III domain from fibronectin (FNfn10) is largely determined by a number of critical hydrogen bonds in the peripheral strands. Interestingly, the simulations predict that lowering the pH from 7 to approximately 4.7 will increase the mechanical stability of FNfn10 significantly (by approximately 33 %) due to the protonation of a few key acidic residues in the A and B strands. To test this simulation prediction, we used single-molecule atomic force microscopy (AFM) to investigate the mechanical stability of FNfn10 at neutral pH and at lower pH where these key residues have been shown to be protonated. Our AFM experimental results show no difference in the mechanical stability of FNfn10 at these different pH values. These results suggest that some simulations may overestimate the role played by electrostatic interactions in determining the mechanical stability of proteins.
Ng
Sean P
SP
Cambridge University Department of Chemistry, MRC Centre for Protein Engineering, Lensfield Rd, Cambridge, CB2 1EW, UK.
Clarke
Jane
J
D020816:Amino Acid Motifs; D003198:Computer Simulation; D005353:Fibronectins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D017510:Protein Folding; D017434:Protein Structure, Tertiary
D005353:Fibronectins