Simulated-annealing real-space refinement as a tool in model building

Methods have been developed that further automate the building of macromolecular structures into electron-density maps. The software supports molecular-dynamics real-space refinement of an atomic model to local regions of a map within the context of a popular molecular-modeling program, O [Jones et al. (1991), Acta Cryst. A47, 110-119]. It is implemented as a module to the CNS refinement package [Brunger et al. (1998), Acta Cryst. D54, 905-921], controlled by a graphical user interface and commands executed directly through the molecular-graphics package. The method is illustrated with examples of the building and rebuilding of protein and nucleic acid models in which laborious manual adjustments are avoided. The resulting models show improved convergence during subsequent reciprocal-space refinement. The novel feature of the RSRef2000 software is the combination of simulated-annealing optimization with local real-space refinement, allowing several local minima to be explored quickly and automatically within the context of interactive model building.