9T6P

Higher-dose (283.1 kGy) structure of copper-containing nitrite reductase from Achromobacter cycloclastes at room temperature

Summary for 9T6P

• Entry DOI: 10.2210/pdb9t6p/pdb
• Descriptor: Copper-containing nitrite reductase, COPPER (II) ION, SULFATE ION, ... (4 entities in total)
• Functional Keywords: nir, cu-nir, radiation damage, nitrite metabolism, room-temperature crystallography, metalloproteins, oxidoreductase
• Biological source: Achromobacter cycloclastes
• Total number of polymer chains: 1
• Total formula weight: 41041.34

Authors

Caramello, N.,Rose, S.L.,Engilberge, S.,Royant, A. (deposition date: 2025-11-07, release date: 2026-03-04, Last modification date: 2026-03-11)

Primary citation

Caramello, N.,Rose, S.L.,Mathieu, E.,Petit, L.,Tews, I.,Engilberge, S.,Royant, A.
Coupled on-line in crystallo UV-Vis absorption spectroscopy and X-ray crystallography to compare specific radiation damage in metal-containing proteins at room versus cryogenic temperature.
Acta Crystallogr D Struct Biol, 82:187-198, 2026

PubMed Abstract: Specific radiation damage (SRD) to proteins is a pertinent issue discovered during the development of cryo-crystallography at synchrotrons, often affecting the macromolecular active site and thus complicating the understanding of mechanistic insights from structural analysis. For proteins with a spectroscopic signature in the visible light spectrum, in crystallo UV-Vis absorption spectroscopy has regularly been used to estimate the dose scale of specific damage build-up and to develop diffraction data-collection strategies to mitigate its effects. Using a coupled spectroscopic and crystallographic approach, here we show that for two metal-containing proteins the structural response to X-ray-induced reduction of metals in their active site is markedly different at room temperature than at cryogenic temperature. This suggests that the use of controlled specific radiation damage to mimic and study a physiological redox transition in a metal-containing protein by X-ray crystallography should preferably be performed at room temperature rather than at cryogenic temperature.

PubMed: 41641581
DOI: 10.1107/S2059798326000690

Experimental method

X-RAY DIFFRACTION (2.008 Å)