Criteria that discriminate between native proteins and incorrectly folded models

Various theoretical concepts, such as free energy potentials, electrostatic interaction potentials, atomic packing, solvent-exposed surface, and surface charge distribution between native proteins and misfolded protein models. Misfolded models were constructed by introducing incorrect side chains onto polypeptide backbones: side chains of the α-helical hemerythrin were modeled on the β-sheeted backbone of immunoglobulin VL domain, whereas those of the VL domain were similarly modeled on he hemerythrin backbone. CONGEN, a conformational space sampling program, was used to construct the side chains, in contrast to the previous work,1 where incorrect side chains were modeled in all trans conformations. Capability of the conformational search procedure to reproduce native conformations was gauged first by rebuilding (the correct) side chains in hemerythrin and the VL domain: constructs with r.m.s differences from the x-ray side chains 2.2–2.4 A were produced, and many calculated conformations matched the native ones quite well. Incorrectly folded models were then constructed by the same conformational protocol applied to incorrect amino acid sequences. All CONGEN constructs, both correctly and incorrectly folded, were characterized by exceptionally small molecular surfaces and low potential energies. Surface charge density, atomic packing, and Coulomb formula-based electrostatic interactions of the misfolded structures and the correctly folded proteins were similar, and therefore of little criteria clearly favored the native structures over the misfolded ones: (1) solvent-exposed side-chain nonpolar surface, (2) number of buried ionizable groups, and (3) empirical free energy functions that incorporate solvent effects.