8RAF

Crystal structure of D-amino acid transaminase from Haliscomenobacter hydrossis point mutant R90I (holo form)

8RAF の概要

• エントリーDOI: 10.2210/pdb8raf/pdb
• 分子名称: Aminotransferase class IV, PYRIDOXAL-5'-PHOSPHATE (3 entities in total)
• 機能のキーワード: daat, point mutant, aminotransferase, transaminase, haliscomenobacter hydrossis, transferase
• 由来する生物種: Haliscomenobacter hydrossis DSM 1100
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 32524.91

構造登録者

Matyuta, I.O.,Bakunova, A.K.,Minyaev, M.E.,Popov, V.O.,Bezsudnova, E.Y.,Boyko, K.M. (登録日: 2023-12-01, 公開日: 2023-12-27, 最終更新日: 2024-05-08)

主引用文献

Bakunova, A.K.,Matyuta, I.O.,Minyaev, M.E.,Isaikina, T.Y.,Boyko, K.M.,Popov, V.O.,Bezsudnova, E.Y.
Multifunctionality of arginine residues in the active sites of non-canonical d-amino acid transaminases.
Arch.Biochem.Biophys., 756:110011-110011, 2024

PubMed Abstract: Structure-function relationships are key to understanding enzyme mechanisms, controlling enzyme activities, and designing biocatalysts. Here, we investigate the functions of arginine residues in the active sites of pyridoxal-5'-phosphate (PLP)-dependent non-canonical d-amino acid transaminases, focusing on the analysis of a transaminase from Haliscomenobacter hydrossis. Our results show that the tandem of arginine residues R28* and R90, which form the conserved R-[RK] motif in non-canonical d-amino acid transaminases, not only facilitates effective substrate binding but also regulates the catalytic properties of PLP. Non-covalent interactions between residues R28*, R90, and Y147 strengthen the hydrogen bond between Y147 and PLP, thereby maintaining the reactivity of the cofactor. Next, the R90 residue contributes to the stability of the holoenzyme. Finally, the R90I substitution induces structural changes that lead to substrate promiscuity, as evidenced by the effective binding of substrates with and without the α-carboxylate group. This study sheds light on the structural determinants of the activity of non-canonical d-amino acid transaminases. Understanding the structural basis of the active site plasticity in the non-canonical transaminase from H. hydrossis, which is characterized by effective conversion of d-amino acids and α-keto acids, may help to tailor it for industrial applications.

PubMed: 38649133
DOI: 10.1016/j.abb.2024.110011

実験手法

X-RAY DIFFRACTION (2 Å)