9AXJ

Cystathionine gamma lyase from Thermobifida fusca in an amino crotonate form

9AXJ の概要

• エントリーDOI: 10.2210/pdb9axj/pdb
• 分子名称: Cystathionine gamma-synthase, (2E)-2-{[(1E)-{3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl}methylidene]amino}but-2-enoic acid, DI(HYDROXYETHYL)ETHER, ... (6 entities in total)
• 機能のキーワード: plp, amino crotonate, lyase
• 由来する生物種: Thermobifida fusca
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 42314.90

構造登録者

Perkins, L.J.,Zmich, A.P.,Bingman, C.A.,Buller, A.R. (登録日: 2024-03-06, 公開日: 2025-01-15)

主引用文献

Zmich, A.,Perkins, L.J.,Bingman, C.,Buller, A.R.
Elucidation of the stereochemical mechanism of cystathionine gamma-lyase reveals how substrate specificity constrains catalysis.
Acs Catalysis, 14:11196-11204, 2024

PubMed Abstract: Pyridoxal phosphate (PLP)-dependent enzymes play essential roles in metabolism and have found applications for organic synthesis and as enzyme therapeutics. The vinylglycine ketimine (VGK) subfamily hosts a growing set of enzymes that play diverse roles in primary and secondary metabolism. However, the molecular determinates of substrate specificity and the complex acid-base chemistry that enables VGK catalysis remain enigmatic. We use a recently discovered amino acid γ-lyase as a model system to probe catalysis in this enzyme family. We discovered that two stereochemically distinct proton transfer pathways occur. Combined kinetic and spectroscopic analysis revealed that progression through the catalytic cycle is correlated with the presence of an H-bond donor after Cγ of an amino acid substrate, suggesting substrate binding is kinetically coupled to a conformational change. High-resolution X-ray crystallography shows that cystathionine-γ-lyases generate an -trans intermediate and that this geometry is likely conserved throughout the VGK family. An H-bond acceptor in the active site templates substrate binding but does so by pre-organizing substrates from catalytically productive orientations. Mutagenesis eliminates this pre-organization, such that there is a relaxation of the substrate specificity, but an increase in for diverse substrates. We exploit this information to perform preparative scale α,β,β-tri-deuteration of polar amino acids. Together, these data untangle a complex mode of substrate specificity and provide a foundation for the future study and applications of VGK enzymes.

PubMed: 39391268
DOI: 10.1021/acscatal.4c02281

実験手法

X-RAY DIFFRACTION (1.5 Å)