Mix-and-extrude: high-viscosity sample injection towards time-resolved protein crystallography.
3D-printed nozzles
X-ray free-electron lasers
membrane proteins
mix-and-extrude
time-resolved serial crystallography
Journal
Journal of applied crystallography
ISSN: 0021-8898
Titre abrégé: J Appl Crystallogr
Pays: United States
ID NLM: 9876190
Informations de publication
Date de publication:
01 Aug 2023
01 Aug 2023
Historique:
received:
23
11
2022
accepted:
21
05
2023
medline:
9
8
2023
pubmed:
9
8
2023
entrez:
9
8
2023
Statut:
epublish
Résumé
Time-resolved crystallography enables the visualization of protein molecular motion during a reaction. Although light is often used to initiate reactions in time-resolved crystallography, only a small number of proteins can be activated by light. However, many biological reactions can be triggered by the interaction between proteins and ligands. The sample delivery method presented here uses a mix-and-extrude approach based on 3D-printed microchannels in conjunction with a micronozzle. The diffusive mixing enables the study of the dynamics of samples in viscous media. The device design allows mixing of the ligands and protein crystals in 2 to 20 s. The device characterization using a model system (fluorescence quenching of iq-mEmerald proteins by copper ions) demonstrated that ligand and protein crystals, each within lipidic cubic phase, can be mixed efficiently. The potential of this approach for time-resolved membrane protein crystallography to support the development of new drugs is discussed.
Identifiants
pubmed: 37555221
doi: 10.1107/S1600576723004405
pii: S1600576723004405
pmc: PMC10405586
doi:
Types de publication
Journal Article
Langues
eng
Pagination
1038-1045Informations de copyright
© Mohammad Vakili et al. 2023.
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