RESUMEN
Currently, one of the most serious problems in protein-folding simulations for de novo structure prediction is conformational sampling of medium-to-large proteins. In vivo, folding of these proteins is mediated by molecular chaperones. Inspired by the functions of chaperonins, we designed a simple chaperonin-like simulation protocol within the framework of the standard fragment assembly method: in our protocol, the strength of the hydrophobic interaction is periodically modulated to help the protein escape from misfolded structures. We tested this protocol for 38 proteins and found that, using a certain defined criterion of success, our method could successfully predict the native structures of 14 targets, whereas only those of 10 targets were successfully predicted using the standard protocol. In particular, for non-alpha-helical proteins, our method yielded significantly better predictions than the standard approach. This chaperonin-inspired protocol that enhanced de novo structure prediction using folding simulations may, in turn, provide new insights into the working principles underlying the chaperonin system.
Asunto(s)
Chaperoninas/química , Chaperoninas/ultraestructura , Modelos Químicos , Modelos Moleculares , Pliegue de Proteína , Proteínas/química , Proteínas/ultraestructura , Simulación por Computador , Conformación ProteicaRESUMEN
Predicting protein tertiary structure by folding-like simulations is one of the most stringent tests of how much we understand the principle of protein folding. Currently, the most successful method for folding-based structure prediction is the fragment assembly (FA) method. Here, we address why the FA method is so successful and its lesson for the folding problem. To do so, using the FA method, we designed a structure prediction test of "chimera proteins." In the chimera proteins, local structural preference is specific to the target sequences, whereas nonlocal interactions are only sequence-independent compaction forces. We find that these chimera proteins can find the native folds of the intact sequences with high probability indicating dominant roles of the local interactions. We further explore roles of local structural preference by exact calculation of the HP lattice model of proteins. From these results, we suggest principles of protein folding: For small proteins, compact structures that are fully compatible with local structural preference are few, one of which is the native fold. These local biases shape up the funnel-like energy landscape.
Asunto(s)
Modelos Químicos , Pliegue de Proteína , Proteínas/química , Proteínas/metabolismo , Simulación por Computador , Modelos Moleculares , Unión Proteica , Desnaturalización Proteica , Estructura Terciaria de ProteínaRESUMEN
Predicting protein tertiary structures by in silico folding is still very difficult for proteins that have new folds. Here, we developed a coarse-grained energy function, SimFold, for de novo structure prediction, performed a benchmark test of prediction with fragment assembly simulations for 38 test proteins, and proposed consensus prediction with Rosetta. The SimFold energy consists of many terms that take into account solvent-induced effects on the basis of physicochemical consideration. In the benchmark test, SimFold succeeded in predicting native structures within 6.5 A for 12 of 38 proteins; this success rate was the same as that by the publicly available version of Rosetta (ab initio version 1.2) run with default parameters. We investigated which energy terms in SimFold contribute to structure prediction performance, finding that the hydrophobic interaction is the most crucial for the prediction, whereas other sequence-specific terms have weak but positive roles. In the benchmark, well-predicted proteins by SimFold and by Rosetta were not the same for 5 of 12 proteins, which led us to introduce consensus prediction. With combined decoys, we succeeded in prediction for 16 proteins, four more than SimFold or Rosetta separately. For each of 38 proteins, structural ensembles generated by SimFold and by Rosetta were qualitatively compared by mapping sampled structural space onto two dimensions. For proteins of which one of the two methods succeeded and the other failed in prediction, the former had a less scattered ensemble located around the native. For proteins of which both methods succeeded in prediction, often two ensembles were mixed up.
Asunto(s)
Proteínas/química , Alanina , Secuencia de Aminoácidos , Simulación por Computador , Secuencia de Consenso , Enlace de Hidrógeno , Modelos Teóricos , Potenciometría , Pliegue de Proteína , Estructura Terciaria de Proteína , TermodinámicaRESUMEN
Protein-folding mechanisms of two small globular proteins, IgG binding domain of protein G and alpha spectrin SH3 domain are investigated via Brownian dynamics simulations with a model made of coarse-grained physical energy functions responsible for sequence-specific interactions and weak Go-like energies. The folding pathways of alpha spectrin SH3 are known to be mainly controlled by the native topology, while protein G folding is anticipated to be more sensitive to the sequence-specific effects than native topology. We found in the folding of protein G that the C terminal beta hairpin is formed earlier and is rigid, once ordered, in the presence of an intact C terminal turn. The alpha helix is found to exhibit repeated partial formations/deformations during folding and to be stabilized via the tertiary contact with preformed beta sheets. This predicted scenario is fully consistent with experimental phi value data. Moreover, we found that the folding route is critically affected when the hydrophobic interaction is excluded from physical energy terms, suggesting that the hydrophobicity critically contributes to the folding propensity of protein G. For the folding of alpha spectrin SH3, we found that the distal beta hairpin and diverging turn are parts formed early, fully in harmony with previous results of simple Go-like and experimental analysis, supporting that the folding route of SH3 domain is robust and coded by the native topology. The hybrid method provides useful tools for analyzing roles of physical interactions in determining folding mechanisms.
Asunto(s)
Modelos Moleculares , Conformación Proteica , Proteínas Bacterianas/química , Simulación por Computador , Inmunoglobulina G/metabolismo , Pliegue de Proteína , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Espectrina/química , Dominios Homologos srcRESUMEN
We optimize a physical energy function for proteins with the use of the available structural database and perform three benchmark tests of the performance: (1) recognition of native structures in the background of predefined decoy sets of Levitt, (2) de novo structure prediction using fragment assembly sampling, and (3) molecular dynamics simulations. The energy parameter optimization is based on the energy landscape theory and uses a Monte Carlo search to find a set of parameters that seeks the largest ratio deltaE(s)/DeltaE for all proteins in a training set simultaneously. Here, deltaE(s) is the stability gap between the native and the average in the denatured states and DeltaE is the energy fluctuation among these states. Some of the energy parameters optimized are found to show significant correlation with experimentally observed quantities: (1) In the recognition test, the optimized function assigns the lowest energy to either the native or a near-native structure among many decoy structures for all the proteins studied. (2) Structure prediction with the fragment assembly sampling gives structure models with root mean square deviation less than 6 A in one of the top five cluster centers for five of six proteins studied. (3) Structure prediction using molecular dynamics simulation gives poorer performance, implying the importance of having a more precise description of local structures. The physical energy function solely inferred from a structural database neither utilizes sequence information from the family of the target nor the outcome of the secondary structure prediction but can produce the correct native fold for many small proteins.