Design of a hyperstable protein by rational consideration of unfolded state interactions

Journal of the American Chemical Society
Burcu AnilDaniel P Raleigh

Abstract

Stabilization of proteins is a long-sought objective. Targeting the unfolded state interactions of a protein is not a method used for this purpose, although many proteins are known to contain such interactions. The N-terminal domain of ribosomal protein L9 (NTL9) has a lysine residue at position 12, which makes strong non-native interactions in the unfolded state. Substitution of a d-alanine for G34 in NTL9 is known to stabilize the protein by reducing the entropy of the unfolded state. Here we combine these two mutations to design a hyperstable protein. The structure of the variant is the same as that of wild-type as judged by 2D NMR. The variant is hyperstable as judged by denaturation experiments, where complete thermal unfolding of the protein does not occur in native buffer.

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Citations

Oct 10, 2008·Journal of the American Chemical Society·Brandon L Kier, Niels H Andersen
Mar 28, 2009·Journal of the Royal Society, Interface·María Suárez, Alfonso Jaramillo
Jan 16, 2016·The Journal of Physical Chemistry. B·Guangfeng Zhou, Vincent Alvin Voelz
Nov 14, 2008·Organic & Biomolecular Chemistry·D Victoria WilliamsNiels H Andersen
Jan 25, 2007·Molecular BioSystems·Bruce E Bowler
Jul 7, 2020·Protein Engineering, Design & Selection : PEDS·Joshua T AtkinsonJonathan J Silberg
Nov 20, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Mahesh Narayan
Jun 26, 2008·Journal of the American Chemical Society·Henry S Ashbaugh, Harold W Hatch
Sep 29, 2007·Archives of Biochemistry and Biophysics·Jae-Hyun ChoDaniel P Raleigh

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