9CGF

Human DJ-1, 20 sec mixing with methylglyoxal, pink beam time-resolved serial crystallography, CrystFEL processed

Summary for 9CGF

• Entry DOI: 10.2210/pdb9cgf/pdb
• Related: 9CEI 9CFI 9CFM 9CFO 9CFQ 9CGA 9CGB 9CGD 9CGE
• Descriptor: Protein deglycase DJ-1, 1-hydroxypropan-2-one (3 entities in total)
• Functional Keywords: glutathione-independent glyoxalase, mix-and-inject serial crystallography, laue diffraction, hydrolase
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 20273.41

Authors

Zielinski, K.,Dolamore, C.,Dalton, K.,Meisburger, S.,Smith, N.,Termini, J.,Henning, R.,Srajer, V.,Hekstra, D.,Pollack, L.,Wilson, M.A. (deposition date: 2024-06-28, release date: 2025-03-12)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.06 Å)