The prediction of protein three-dimensional structure from amino acid sequence has been a grand challenge problem in computational biophysics for decades, owing to its intrinsic scientific interest and also to the many potential applications for robust protein structure prediction algorithms, from genome interpretation to protein function prediction. More recently, the inverse problem — designing an amino acid sequence that will fold into a specified three-dimensional structure — has attracted growing attention as a potential route to the rational engineering of proteins with functions useful in biotechnology and medicine. Methods for the prediction and design of protein structures have advanced dramatically in the past decade. Increases in computing power and the rapid growth in protein sequence and structure databases have fuelled the development of new data-intensive and computationally demanding approaches for structure prediction. New algorithms for designing protein folds and protein–protein interfaces have been used to engineer novel high-order assemblies and to design from scratch fluorescent proteins with novel or enhanced properties, as well as signalling proteins with therapeutic potential. In this Review, we describe current approaches for protein structure prediction and design and highlight a selection of the successful applications they have enabled.
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The authors apologize to the scientists whose important work could not be cited in this Review owing to space constraints. This work was supported by NIH grants R01GM117968 and R35GM131923 to B.K. and R01GM121487 and R01GM123378 to P.B.
The authors declare no competing interests.
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CASP protein structure prediction experiment: http://predictioncenter.org/casp13
- Protein energy functions
Functions that correspond to a mathematical model of the molecular forces that determine protein structures and interactions. The choice of an energy function defines a map from structures onto energy values, referred to as an energy landscape, which can guide structure prediction and design simulations. Typical protein energy functions are linear combinations of multiple terms, each term capturing a distinct energetic contribution (van der Waals interactions, electrostatics, desolvation), with the weights and atomic parameters for these terms chosen by a parameterization procedure that seeks to optimize the agreement between the quantities predicted from the energy function and the corresponding values derived from experiments or from quantum chemistry calculations on small chemical systems.
- Deep learning
A form of machine learning that employs artificial neural networks with many internal-processing layers to recognize patterns in large and complex datasets, such as visual images and written and spoken language.
- Van der Waals interactions
Inter-atomic or inter-molecular interactions that are individually weak (much weaker than covalent or ionic bonds) and relatively short-ranged.
A discrete set of conformations frequently adopted by amino acid side chains.
- Degrees of freedom
The free parameters in a system that determine its structure and, hence, its energy. They can be continuous, such as a real-valued backbone torsion angle or atomic position, or discrete (permitting only a finite number of alternatives). Owing to strong torsional preferences, side-chain conformations can be successfully modelled using a discrete set of rotamers, identified by analysis of the structural database.
- Phi angle
A torsion angle (or dihedral angle) that describes rotation about the bond that connects the backbone nitrogen and the backbone Cα carbon of an amino acid in a polypeptide chain. It is one of the two primary degrees of freedom (along with the psi angle) per amino acid residue that proteins use to adopt alternative conformations.
- Psi angle
A torsion angle (or dihedral angle) that describes rotation about the bond that connects the backbone Cα carbon and backbone carbonyl carbon of an amino acid in a polypeptide chain. It is one of the two primary degrees of freedom (along with the phi angle) per amino acid residue that proteins use to adopt alternative conformations.
- Chi1 angle
A torsion angle (or dihedral angle) in amino acid side chains that is numbered on the basis of the proximity in chemical connectivity of the bond to the protein backbone. Chi1 refers to rotation about the bond closest to the backbone, chi2 is the next closest position and so on. Some amino acids, such as alanine and glycine, have no rotatable bonds or torsion angles, while others, such as lysine and arginine, have up to four.
A software package for the prediction and design of protein structures and interactions that implements a wide range of backbone and side-chain conformational sampling algorithms and sequence optimization methods.
- Dead-end elimination
An algorithm for side-chain rotamer optimization that functions by eliminating rotamers that are not compatible with adopting the sequence with the lowest possible energy.
- Mean-field optimization
A protocol for designing sequences that assigns a probability to observing each amino acid at each sequence position in the protein and calculates an average (mean-field) energy for the protein based on the assigned probabilities. The probabilities are then adjusted to lower the mean-field energy of the protein.
- Simulated annealing
A probabilistic approach for identifying low-energy sequences that accepts or rejects sequence changes on the basis of the calculated change in the energy of the protein when a sequence change is made and the temperature of the modelled system. If a change lowers the energy of the protein, it is automatically accepted; if it raises the energy of the system, it is accepted with some probability that depends on how much the energy has increased (a bigger increase in energy is less likely to be accepted) and the current temperature (at higher temperatures it is more likely to accept changes that raise the energy of the system). The temperature is lowered as the simulation progresses, to identify low-energy sequences.
- Genetic algorithms
A sequence optimization protocol that repeatedly modifies a population of sequences by applying rounds of energy-based selection. The energy of a sequence is calculated by modelling it in the desired protein conformation. Lower-energy sequences are more likely to progress to the next generation. Before each round of selection, the previous winning sequences are recombined with each other and small numbers of mutations are incorporated into the sequences, to search for lower-energy sequences.
- Multi-state design algorithms
An approach to designing sequences that satisfy multiple constraints simultaneously. For instance, such algorithms can be used to find protein sequences that are predicted to bind ligand X but not ligand Y. Alternatively, they can be repurposed to find sequences that are simultaneously good at binding both ligand X and ligand Y.
- Yeast surface display
An experimental approach for probing a large library of protein sequences (up to tens of millions) for binding to another molecule. In the final yeast library, each yeast cell contains the DNA for one member of the protein library, and this protein is expressed as a fusion protein that presents the protein on the outside of the cell. The cells are mixed with the target molecule, which has been labelled with a fluorescent dye, and then fluorescence-activated cell sorting is used to identify the cells that contain a designed protein that binds the target protein. DNA sequencing is used to identify the designs that passed selection.
- BCL-2 protein family
A family of structurally related proteins that interact with each other to induce or repress apoptosis.
- DNA origami
A term describing approaches that use the high sequence specificity of DNA interactions to design DNA sequences that will fold into complex and predictable two- and three-dimensional shapes.
- Major histocompatibility complexes
(MHCs). A set of cell surface proteins that bind to antigens from foreign pathogens and present them for recognition by other proteins and cells from the immune system. They are a key component of the acquired immune system.
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Kuhlman, B., Bradley, P. Advances in protein structure prediction and design. Nat Rev Mol Cell Biol 20, 681–697 (2019). https://doi.org/10.1038/s41580-019-0163-x
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