Homology Modeling
From DrugPedia: A Wikipedia for Drug discovery
Homology Modeling is a method of prediction of protein 3D structure from the amino acid sequence.This is also known as Comparative modeling.This process in based on the selection of templete molecule whose 3D structure is known.
Homology modeling can produce high-quality structural models when the target and template are closely related, which has inspired the formation of a structural genomics consortium dedicated to the production of representative experimental structures for all classes of protein folds.[3] The chief inaccuracies in homology modeling, which worsen with lower sequence identity, derive from errors in the initial sequence alignment and from improper template selection.[4] Like other methods of structure prediction, current practice in homology modeling is assessed in a biannual large-scale experiment known as the Critical Assessment of Techniques for Protein Structure Prediction, or CASP.
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[edit] Steps in homology modeling
- Selection of Templete molecule
- Alignment of Templete with Target
- Model Generation
- Model Assesment
[edit] Template Selection
If the percentage sequence identity between the sequence of interest and a protein with known structure is high enough (more than 25 or 30 %) simple database search programs like FASTA or BLAST are clearly adequate to detect the homology.
[edit] Templete Alignment
A critical step in the development of a homology model is the alignment of the unknown sequence with the homologues. Factors to be considered when performing an alignment are
(1) which algorithm to use for sequence alignment (2) which scoring method to apply (3) whether and how to assign gap penalties
[edit] Model Generation
Given a template and an alignment, the information contained therein must be used to generate a three-dimensional structural model of the target, represented as a set of Cartesian coordinates for each atom in the protein. Three major classes of model generation methods have been proposed.
[edit] Fragment assembly
The original method of homology modeling relied on the assembly of a complete model from conserved structural fragments identified in closely related solved structures. For example, a modeling study of serine proteases in mammals identified a sharp distinction between "core" structural regions conserved in all experimental structures in the class, and variable regions typically located in the loops where the majority of the sequence differences were localized. Thus unsolved proteins could be modeled by first constructing the conserved core and then substituting variable regions from other proteins in the set of solved structures. Current implementations of this method differ mainly in the way they deal with regions that are not conserved or that lack a template.
[edit] Segment matching
The segment-matching method divides the target into a series of short segments, each of which is matched to its own template fitted from the Protein Data Bank. Thus, sequence alignment is done over segments rather than over the entire protein. Selection of the template for each segment is based on sequence similarity, comparisons of alpha carbon coordinates, and predicted steric conflicts arising from the van der Waals radii of the divergent atoms between target and template.
[edit] Satisfaction of spatial restraints
The most common current homology modeling method takes its inspiration from calculations required to construct a three-dimensional structure from data generated by NMR spectroscopy. One or more target-template alignments are used to construct a set of geometrical criteria that are then converted to probability density functions for each restraint. Restraints applied to the main protein internal coordinates - protein backbone distances and dihedral angles - serve as the basis for a global optimization procedure that originally used conjugate gradient energy minimization to iteratively refine the positions of all heavy atoms in the protein.
This method had been dramatically expanded to apply specifically to loop modeling, which can be extremely difficult due to the high flexibility of loops in proteins in aqueous solution. A more recent expansion applies the spatial-restraint model to electron density maps derived from cryoelectron microscopy studies, which provide low-resolution information that is not usually itself sufficient to generate atomic-resolution structural models. To address the problem of inaccuracies in initial target-template sequence alignment, an iterative procedure has also been introduced to refine the alignment on the basis of the initial structural fit. The most commonly user software in spatial restraint-based modeling is MODELLER and a database called ModBase has been established for reliable models generated with it.
[edit] Model Assesment
Assessment of homology models without reference to the true target structure is usually performed with two methods: statistical potentials or physics-based energy calculations. Both methods produce an estimate of the energy (or an energy-like analog) for the model or models being assessed; independent criteria are needed to determine acceptable cutoffs. Neither of the two methods correlates exceptionally well with true structural accuracy, especially on protein types underrepresented in the PDB, such as membrane proteins.
Statistical potentials are empirical methods based on observed residue-residue contact frequencies among proteins of known structure in the PDB. They assign a probability or energy score to each possible pairwise interaction between amino acids and combine these pairwise interaction scores into a single score for the entire model. Some such methods can also produce a residue-by-residue assessment that identifies poorly scoring regions within the model, though the model may have a reasonable score overall. These methods emphasize the hydrophobic core and solvent-exposed polar amino acids often present in globular proteins. Examples of popular statistical potentials include Prosa and DOPE. Statistical potentials are more computationally efficient than energy calculations.
Physics-based energy calculations aim to capture the interatomic interactions that are physically responsible for protein stability in solution, especially van der Waals and electrostatic interactions. These calculations are performed using a molecular mechanics force field; proteins are normally too large even for semi-empirical quantum mechanics-based calculations. The use of these methods is based on the energy landscape hypothesis of protein folding, which predicts that a protein's native state is also its energy minimum. Such methods usually employ implicit solvation, which provides a continuous approximation of a solvent bath for a single protein molecule without necessitating the explicit representation of individual solvent molecules. A force field specifically constructed for model assessment is known as the Effective Force Field (EFF) and is based on atomic parameters from CHARMM.
A very extensive model validation report can be obtained using the Radboud Universiteit Nijmegen "What Check" software which is one option of the Radboud Universiteit Nijmegen "What If" software package; it produces a many page document with extensive analyses of nearly 200 scientific and administrative aspects of the model. "What Check" is available as a free server; it can also be used to validate experimentally determined structures of macromolecules.
One newer method for model assessment relies on machine learning techniques such as neural nets, which may be trained to assess the structure directly or to form a consensus among multiple statistical and energy-based methods. Very recent results using support vector machine regression on a jury of more traditional assessment methods outperformed common statistical, energy-based, and machine learning methods.
[edit] Structural comparison methods
The assessment of homology models' accuracy is straightforward when the experimental structure is known. The most common method of comparing two protein structures uses the root-mean-square deviation (RMSD) metric to measure the mean distance between the corresponding atoms in the two structures after they have been superimposed. However, RMSD does underestimate the accuracy of models in which the core is essentially correctly modeled, but some flexible loop regions are inaccurate. A method introduced for the modeling assessment experiment CASP is known as the global distance test (GDT) and measures the total number of atoms whose distance from the model to the experimental structure lies under a certain distance cutoff. Both methods can be used for any subset of atoms in the structure, but are often applied to only the alpha carbon or protein backbone atoms to minimize the noise created by poorly modeled side chain rotameric states, which most modeling methods are not optimized to predict.
