PDBsum entry 6y9f

Hydrolase

PDB id: 6y9f

Contents

Protein chain

288 a.a.

Ligands

NHE ×2

Waters ×272

References listed in PDB file

Key reference

• Title: Structures of hyperstable ancestral haloalkane dehalogenases show restricted conformational dynamics.
• Authors: P.Babkova, Z.Dunajova, R.Chaloupkova, J.Damborsky, D.Bednar, M.Marek.
• Ref.: Comput Struct Biotechnol J, 2020, 18, 1497-1508. [DOI no: 10.1016/j.csbj.2020.06.021]
• PubMed id: 32637047

Abstract

Ancestral sequence reconstruction is a powerful method for inferring ancestors of modern enzymes and for studying structure-function relationships of enzymes. We have previously applied this approach to haloalkane dehalogenases (HLDs) from the subfamily HLD-II and obtained thermodynamically highly stabilized enzymes (ΔT_m up to 24 °C), showing improved catalytic properties. Here we combined crystallographic structural analysis and computational molecular dynamics simulations to gain insight into the mechanisms by which ancestral HLDs became more robust enzymes with novel catalytic properties. Reconstructed ancestors exhibited similar structure topology as their descendants with the exception of a few loop deviations. Strikingly, molecular dynamics simulations revealed restricted conformational dynamics of ancestral enzymes, which prefer a single state, in contrast to modern enzymes adopting two different conformational states. The restricted dynamics can potentially be linked to their exceptional stabilization. The study provides molecular insights into protein stabilization due to ancestral sequence reconstruction, which is becoming a widely used approach for obtaining robust protein catalysts.