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Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip
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Biochemie
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JoVE Journal Biochemie
Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip

Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip

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10:45 min

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March 20, 2021

DOI:

10.3791/61972-v

DOI:
10.3791/61972-v

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10:45 min
March 20, 2021

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1Université de Strasbourg, Architecture et Réactivité de l’ARN, UPR 9002, CNRS,Institut de Biologie Moléculaire et Cellulaire, 2Biochemistry and Molecular Biology, Institute for Biochemistry,Leipzig University, 3Paul Scherrer Institute, Swiss Light Source, 4European XFEL GmbH

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A versatile microfluidic device is described that enables the crystallization of an enzyme using the counter-diffusion method, the introduction of a substrate in the crystals by soaking, and the 3D structure determination of the enzyme:substrate complex by a serial analysis of crystals inside the chip at room temperature.

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CrystallizationStructural DeterminationEnzyme-substrate ComplexSerial CrystallographyMicrofluidic ChipCounter-diffusionIn Situ X-ray DiffractionBiomoleculeCrystal GrowthRoom Temperature Analysis

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de Wijn, R., Rollet, K., Olieric, V., Hennig, O., Thome, N., Noûs, C., Paulus, C., Lorber, B., Betat, H., Mörl, M., Sauter, C. Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip. J. Vis. Exp. (169), e61972, doi:10.3791/61972 (2021).

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