The production of enzyme microcrystals for time resolved serial crystallography employing free electron laser or synchrotron radiation is a relatively new variation on traditional macromolecular crystallization for conventional single crystal X-ray analysis. While the fundamentals of macromolecular crystal growth are the same, some modifications and special considerations are in order if the objective is to produce uniform size, microcrystals in very large numbers for serial data collection. Presented here are the basic principles of protein crystal growth with particular attention to the approaches best employed to achieve the goal of microcrystals and some novel techniques, as well as old, that may be useful. Also discussed are the advantages of particular precipitants and certain methods of growing protein crystals that might be advantageous for serial data recording.
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http://dx.doi.org/10.1016/bs.mie.2024.10.003 | DOI Listing |
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November 2024
State Key Laboratory of Chemical Resource Engineering, College of Chemical Engineering, Beijing University of Chemical Technology, Beijing, 100029, China.
Methods Enzymol
November 2024
Diamond Light Source, Harwell Science & Innovation Campus, Didcot, United Kingdom; Research Complex at Harwell, Rutherford Appleton Laboratory, Didcot, United Kingdom. Electronic address:
Dynamic structural biology enables studying biological events at the atomic scale from 10's of femtoseconds to a few seconds duration. With the advent of X-ray Free Electron Lasers (XFELs) and 4th generation synchrotrons, serial crystallography is becoming a major player for time-resolved experiments in structural biology. Despite significant progress, challenges such as obtaining sufficient amounts of protein to produce homogeneous microcrystal slurry, remain.
View Article and Find Full Text PDFMethods Enzymol
November 2024
Diamond Light Source Ltd, Harwell Science and Innovation Campus, Didcot, Oxfordshire, United Kingdom; Research Complex at Harwell, Harwell Science and Innovation Campus, Didcot, Oxfordshire, United Kingdom. Electronic address:
Methods Enzymol
November 2024
Graduate School of Science, University of Hyogo, Kouto, Kamigori, Ako, Hyogo, Japan. Electronic address:
Methods Enzymol
November 2024
Department of Molecular Biology and Biochemistry, University of CA Irvine, Irvine, CA, United States; The Scripps Research Institute Florida, Jupiter, FL, United States. Electronic address:
The production of enzyme microcrystals for time resolved serial crystallography employing free electron laser or synchrotron radiation is a relatively new variation on traditional macromolecular crystallization for conventional single crystal X-ray analysis. While the fundamentals of macromolecular crystal growth are the same, some modifications and special considerations are in order if the objective is to produce uniform size, microcrystals in very large numbers for serial data collection. Presented here are the basic principles of protein crystal growth with particular attention to the approaches best employed to achieve the goal of microcrystals and some novel techniques, as well as old, that may be useful.
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