Effects of disulfide bonds on folding behavior and mechanism of the beta-sheet protein tendamistat.
Tendamistat, a small disulfide-bonded beta-sheet protein, and its three single/double-disulfide mutants are investigated by using a modified Gō-like model, aiming to understand the folding mechanism of disulfide-bonded protein as well as the effects of removal of disulfide bond on the folding process. Our simulations show that tendamistat and its two single-disulfide mutants are all two-state folders, consistent with the experimental observations. It is found that the disulfide bonds as well as three hydrogen bonds between the N-terminal loop-0 and strand-6 are of significant importance for the folding of tendamistat. Without these interactions, their two-state behaviors become unstable and the predictions of the model are inconsistent with experiments. In addition, the effect of disulfide bonds on the folding process are studied by comparing the wild-type tendamistat and its two mutants; it is found that the removal of either of the C11-C27 or C45-C73 disulfide bond leads to a large decrease in the thermodynamical stability and loss of structure in the unfolded state, and the effect of the former is stronger than that of the later. These simulation results are in good agreement with experiments and, thus, validate our model. Based on the same model, the detailed folding pathways of the wild-type tendamistat and two mutants are studied, and the effect of disulfide bonds on the folding kinetics are discussed. The obtained results provide a detailed folding picture of these proteins and complement experimental findings. Finally, the folding nuclei predicted to be existent in this protein tendamistat as well as its mutants are firstly identified in this work. The positions of the nucleus are consistent with those argued in experimental studies. Therefore, a nucleation/growth folding mechanism that can explain the two-state folding manner is clearly characterized. Moreover, the effect by the removal of each disulfide bond on the folding thermodynamics and dynamics can also be well interpreted from their influence on the folding nucleus. The implementation of this work indicates that the modified Gō-like model really describes the folding behavior of protein tendamistat and could be used to study the folding of other disulfide-bonded proteins.
Qin
Meng
M
National Laboratory of Solid State Microstructure and Department of Physics, Nanjing University, Nanjing 210093, China.
Zhang
Jian
J
Wang
Wei
W
D055592:Biophysical Phenomena; D001703:Biophysics; D002467:Cell Nucleus / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry; D006358:Hot Temperature; D006860:Hydrogen Bonding; D007700:Kinetics; D008958:Models, Molecular; D015233:Models, Statistical; D009154:Mutation; D010455:Peptides / Q000737:chemistry*; D011336:Probability; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013302:Streptomyces / Q000378:metabolism; D013696:Temperature; D013816:Thermodynamics; D013997:Time Factors
D004220:Disulfides; D004791:Enzyme Inhibitors; D010455:Peptides; C030526:tendamistate