8BHC

K141H and S142H double mutant of hGSTA1-1

8BHC の概要

• エントリーDOI: 10.2210/pdb8bhc/pdb
• 分子名称: Glutathione S-transferase A1 (2 entities in total)
• 機能のキーワード: thermostability, glutathione, detoxification, catalysis, transferase
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 103225.10

構造登録者

Papageorgiou, A.C.,Poudel, N. (登録日: 2022-10-31, 公開日: 2023-03-08, 最終更新日: 2024-11-20)

主引用文献

Chronopoulou, E.G.,Mutabdzija, L.,Poudel, N.,Papageorgiou, A.C.,Labrou, N.E.
A Key Role in Catalysis and Enzyme Thermostability of a Conserved Helix H5 Motif of Human Glutathione Transferase A1-1.
Int J Mol Sci, 24:-, 2023

PubMed Abstract: Glutathione transferases (GSTs) are promiscuous enzymes whose main function is the detoxification of electrophilic compounds. These enzymes are characterized by structural modularity that underpins their exploitation as dynamic scaffolds for engineering enzyme variants, with customized catalytic and structural properties. In the present work, multiple sequence alignment of the alpha class GSTs allowed the identification of three conserved residues (E137, K141, and S142) at α-helix 5 (H5). A motif-directed redesign of the human glutathione transferase A1-1 (hGSTA1-1) was performed through site-directed mutagenesis at these sites, creating two single- and two double-point mutants (E137H, K141H, K141H/S142H, and E137H/K141H). The results showed that all the enzyme variants displayed enhanced catalytic activity compared to the wild-type enzyme hGSTA1-1, while the double mutant hGSTA1-K141H/S142H also showed improved thermal stability. X-ray crystallographic analysis revealed the molecular basis of the effects of double mutations on enzyme stability and catalysis. The biochemical and structural analysis presented here will contribute to a deeper understanding of the structure and function of alpha class GSTs.

PubMed: 36835112
DOI: 10.3390/ijms24043700

実験手法

X-RAY DIFFRACTION (1.56 Å)