9CFI

Human DJ-1, 3 sec mixing with methylglyoxal, pink beam time-resolved serial crystallography

9CFI の概要

• エントリーDOI: 10.2210/pdb9cfi/pdb
• 関連するPDBエントリー: 9CEI
• 分子名称: Protein deglycase DJ-1, 1-hydroxypropan-2-one (3 entities in total)
• 機能のキーワード: glutathione-independent glyoxalase, mix-and-inject serial crystallgraphy, laue diffraction, hydrolase
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 20273.41

構造登録者

Zielinski, K.A.,Dolamore, C.,Dalton, K.,Meisburger, S.P.,Smith, N.,Termini, J.,Henning, R.,Srajer, V.,Hekstra, D.,Pollack, L.,Wilson, M.A. (登録日: 2024-06-27, 公開日: 2025-03-12)

主引用文献

Zielinski, K.A.,Dolamore, C.,Dalton, K.M.,Smith, N.,Termini, J.,Henning, R.,Srajer, V.,Hekstra, D.R.,Pollack, L.,Wilson, M.A.
Resolving DJ-1 Glyoxalase Catalysis Using Mix-and-Inject Serial Crystallography at a Synchrotron.
Biorxiv, 2024

PubMed Abstract: DJ-1 (PARK7) is an intensively studied protein whose cytoprotective activities are dysregulated in multiple diseases. DJ-1 has been reported as having two distinct enzymatic activities in defense against reactive carbonyl species that are difficult to distinguish in conventional biochemical experiments. Here, we establish the mechanism of DJ-1 using a synchrotron-compatible version of mix-and-inject-serial crystallography (MISC), which was previously performed only at XFELs, to directly observe DJ-1 catalysis. We designed and used new diffusive mixers to collect time-resolved Laue diffraction data of DJ-1 catalysis at a pink beam synchrotron beamline. Analysis of structurally similar methylglyoxal-derived intermediates formed through the DJ-1 catalytic cycle shows that the enzyme catalyzes nearly two turnovers in the crystal and defines key aspects of its glyoxalase mechanism. In addition, DJ-1 shows allosteric communication between a distal site at the dimer interface and the active site that changes during catalysis. Our results rule out the widely cited deglycase mechanism for DJ-1 action and provide an explanation for how DJ-1 produces L-lactate with high chiral purity.

PubMed: 39071394
DOI: 10.1101/2024.07.19.604369

実験手法

X-RAY DIFFRACTION (1.77 Å)