A miniature airlock system to aid the cryo-cooling of protein crystals grown under anoxic conditions

Autor: Peter van der Linden, Sylvain Engilberge, Mohamed Atta, Philippe Carpentier
Přispěvatelé: Partnership for Soft Condensed Matter, European Synchrotron Radiation Facility, European Synchroton Radiation Facility [Grenoble] (ESRF), Biocatalyse (BIOCAT), Laboratoire de Chimie et Biologie des Métaux (LCBM - UMR 5249), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Laboratoire de Chimie des Processus Biologiques (LCPB), Collège de France (CdF (institution))-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), ANR-17-EURE-0003,CBH-EUR-GS,CBH-EUR-GS(2017)
Rok vydání: 2022
Předmět:
Zdroj: Journal of Applied Crystallography
Journal of Applied Crystallography, 2022, 55 (5), pp.1212-1218. ⟨10.1107/S1600576722007580⟩
ISSN: 1600-5767
0021-8898
DOI: 10.1107/s1600576722007580
Popis: Crystallographic studies of structures and mechanisms of dioxygen-employing enzymes or oxygen-sensitive proteins usually require these molecules to be prepared and crystallized under anoxic conditions in gloveboxes. Handling, cryo-cooling and extraction of these crystals from gloveboxes for X-ray data collection can be a tricky operation, during which samples might be ruined if they are accidentally exposed to atmospheric oxygen. This work reports the design of a new miniature airlock system and describes the associated method in order to simplify these tasks. The system allows flash-cooling of crystals harvested in a glovebox directly into an external liquid nitrogen container via a miniature airlock while maintaining strict anoxic conditions. The method was assessed using hen egg-white lysozyme crystals soaked in dihydroresorufin (an O2 indicator) and with the reduced tRNA-hydroxylase enzyme MiaE.
Databáze: OpenAIRE
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