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From coiled coils to small globular proteins: Design of a native‐like three‐helix bundle
Author(s) -
Bryson James W.,
Desjarlais John R.,
Handel Tracy M.,
Degrado William F.
Publication year - 1998
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560070617
Subject(s) - helix bundle , antiparallel (mathematics) , circular dichroism , globular protein , crystallography , chemistry , helix (gastropod) , folding (dsp implementation) , protein folding , bundle , nuclear magnetic resonance spectroscopy , protein design , native state , electrostatics , fluorescence spectroscopy , lattice protein , guanidine , spectroscopy , coiled coil , protein structure , fluorescence , stereochemistry , materials science , physics , organic chemistry , biochemistry , biology , snail , ecology , engineering , composite material , quantum mechanics , magnetic field , electrical engineering
A monomolecular native‐like three‐helix bundle has been designed in an iterative process, beginning with a peptide that noncooperatively assembled into an antiparallel three‐helix bundle. Three versions of the protein were designed in which specific interactions were incrementally added. The hydrodynamic and spectroscopic properties of the proteins were examined by size exclusion chromatography, sedimentation equilibrium, fluorescence spectroscopy, and NMR. The thermodynamics of folding were evaluated by monitoring the thermal and guanidine‐induced unfolding transitions using far UV circular dichroism spectroscopy. The attainment of a unique, native‐like state was achieved through the introduction of: (1) helix capping interactions; (2) electrostatic interactions between partially exposed charged residues; (3) a diverse collection of apolar side chains within the hydrophobic core.