4U3H

Crystal structure of FN3con

4U3H の概要

• エントリーDOI: 10.2210/pdb4u3h/pdb
• 分子名称: FN3con (2 entities in total)
• 機能のキーワード: fibronectin type iii, fn3, consensus design, protein stability, de novo protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 10759.84

構造登録者

Porebski, B.T.,McGowan, S.,Buckle, A.M. (登録日: 2014-07-21, 公開日: 2015-02-11, 最終更新日: 2023-09-27)

主引用文献

Porebski, B.T.,Nickson, A.A.,Hoke, D.E.,Hunter, M.R.,Zhu, L.,McGowan, S.,Webb, G.I.,Buckle, A.M.
Structural and dynamic properties that govern the stability of an engineered fibronectin type III domain.
Protein Eng.Des.Sel., 28:67-78, 2015

PubMed Abstract: Consensus protein design is a rapid and reliable technique for the improvement of protein stability, which relies on the use of homologous protein sequences. To enhance the stability of a fibronectin type III (FN3) domain, consensus design was employed using an alignment of 2123 sequences. The resulting FN3 domain, FN3con, has unprecedented stability, with a melting temperature >100°C, a ΔG(D-N) of 15.5 kcal mol(-1) and a greatly reduced unfolding rate compared with wild-type. To determine the underlying molecular basis for stability, an X-ray crystal structure of FN3con was determined to 2.0 Å and compared with other FN3 domains of varying stabilities. The structure of FN3con reveals significantly increased salt bridge interactions that are cooperatively networked, and a highly optimized hydrophobic core. Molecular dynamics simulations of FN3con and comparison structures show the cooperative power of electrostatic and hydrophobic networks in improving FN3con stability. Taken together, our data reveal that FN3con stability does not result from a single mechanism, but rather the combination of several features and the removal of non-conserved, unfavorable interactions. The large number of sequences employed in this study has most likely enhanced the robustness of the consensus design, which is now possible due to the increased sequence availability in the post-genomic era. These studies increase our knowledge of the molecular mechanisms that govern stability and demonstrate the rising potential for enhancing stability via the consensus method.

PubMed: 25691761
DOI: 10.1093/protein/gzv002

実験手法

X-RAY DIFFRACTION (1.98 Å)