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Published June 1, 1998 | public
Journal Article

Design, structure and stability of a hyperthermophilic protein variant


Here we report the use of an objective computer algorithm in the design of a hyperstable variant of the Streptococcal protein Gβ1 domain (Gβ1). The designed seven-fold mutant, Gβ1-c3b4, has a melting temperature in excess of 100 degrees C and an enhancement in thermodynamic stability of 4.3 kcal mol^(-1) at 50 degrees C over the wild-type protein. Gβ1-c3b4 maintains the Gβ1 fold, as determined by nuclear magnetic resonance spectroscopy, and also retains a significant level of binding to human IgG in qualitative comparisons with wild type. The basis of the stability enhancement appears to have multiple components including optimized core packing, increased burial of hydrophobic surface area, more favorable helix dipole interactions, and improvement of secondary structure propensity. The design algorithm is able to model such complex contributions simultaneously using empirical physical/chemical potential functions and a combinatorial optimization algorithm based on the dead-end elimination theorem. Because the design methodology is based on general principles, there is the potential of applying the methodology to the stabilization of other unrelated protein folds.

Additional Information

© 1998 Nature Publishing Group. Received March 20, 1998; accepted May 1, 1998. We thank P. Poon for sedimentation equilibrium measurements and discussions, B. Dahiyat, A. Su, S. Ross, D. Bökenkamp, and C. Sariskyfor technical assistance and discussions, and G. Hathaway for mass spectrometry. Supported by the Howard Hughes Medical Institute (S.L.M.), the Rita Alien Foundation, the Chandler Family Trust, the Booth Ferris Foundation, the David and Lucile Packard Foundation, the Searle Scholars Program and The Chicago Community Trust and an NIH training grant (S.M.M.)

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