title: Increasing protein conformational stability by optimizing beta-turn sequence.
abstract: Protein conformational stability is an important concern in many fields. Here, we describe a strategy for significantly increasing conformational stability by optimizing beta-turn sequence. Proline and glycine residues are statistically preferred at several beta-turn positions, presumably because their unique side-chains contribute favorably to conformational stability in certain beta-turn positions. However, beta-turn sequences often deviate from preferred proline or preferred glycine. Therefore, our strategy involves replacing non-proline and non-glycine beta-turn residues with preferred proline or preferred glycine residues. Here, we develop guidelines for selecting appropriate mutations, and present results for five mutations (S31P, S42G, S48P, T76P, and Q77G) that significantly increase the conformational stability of RNase Sa. The increases in stability ranged from 0.7 kcal/mol to 1.3 kcal/mol. The strategy was successful in overlapping or isolated beta-turns, at buried (up to 50%) or completely exposed sites, and at relatively flexible or inflexible sites. Considering the significant number of beta-turn residues in every globular protein and the frequent deviation of beta-turn sequences from preferred proline and preferred glycine residues, this simple, efficient strategy will be useful for increasing the conformational stability of proteins.
authors.affiliation: Department of Molecular and Cellular Medicine, Texas A&M University System Health Science Center, College Station, TX 77843, USA.
authors.forename: Saul R
authors.initials: SR
authors.lastname: Trevino
authors.forename: Stephanie
authors.initials: S
authors.lastname: Schaefer
authors.forename: J Martin
authors.initials: JM
authors.lastname: Scholtz
authors.forename: C Nick
authors.initials: CN
authors.lastname: Pace
mesh_terms: D005998:Glycine / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D011392:Proline / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D013816:Thermodynamics
chemical_list: D011392:Proline; D005998:Glycine