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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

DOI:
10.3791/61972-v

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10:45 min

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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

Kapitel

  • 00:00Introduction
  • 01:10ChipX Presentation
  • 02:25Loading Biomolecule Samples in ChipX
  • 03:12Loading Crystallization Solutions in ChipX
  • 04:10Growing Crystals in ChipX
  • 05:03Serial Crystallography Analysis in ChipX
  • 08:33Representative Results
  • 09:55Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

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.

Tags

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

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