Optimization of designed armadillo repeat proteins by molecular dynamics simulations and NMR spectroscopy.

ABSTRACT: A multidisciplinary approach based on molecular dynamics (MD) simulations using homology models, NMR spectroscopy, and a variety of biophysical techniques was used to efficiently improve the thermodynamic stability of armadillo repeat proteins (ArmRPs). ArmRPs can form the basis of modular peptide recognition and the ArmRP version on which synthetic libraries are based must be as stable as possible. The 42-residue internal Arm repeats had been designed previously using a sequence-consensus method. Heteronuclear NMR revealed unfavorable interactions present at neutral but absent at high pH. Two lysines per repeat were involved in repulsive interactions, and stability was increased by mutating both to glutamine. Five point mutations in the capping repeats were suggested by the analysis of po

SUBMITTER: Alfarano P 

PROVIDER: S-EPMC3631359 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature