Preparation of the Crystals of [NiFe] Hydrogenase Suitable for the Neutron Structure

DOI:10.15011//jasma.34.340106
Int. J. Microgravity Sci. Appl. 2017p340106
Author
Koji NISHIKAWA and Yoshiki HIGUCHI
Organization
Graduate School of Life Science, University of Hyogo
Abstract
X-ray structure analysis method has been widely utilized to solve the structure of biological macromolecules. It is, however, also well known that the hydrogen atoms and protons, which are important to understand the structure-function relationship of them, are hardly visible in the electron density map due to the small atomic scattering factor of hydrogen. To visualize hydrogen species, neutron structure analysis is the most powerful technique, because the nuclear scattering length of hydrogen is comparable to those of other atoms comprising the protein molecules. Though many x-ray crystal structures of [NiFe] hydrogenases were reported so far, the real proton pathway and reaction mechanism at the Ni-Fe active site are still unclear. In this paper, we report how to prepare the high-quality large single crystals by using an improved crystallization phase diagram for [NiFe] hydrogenase.
Keyword(s)
[NiFe] Hydrogenase, Neutron structure analysis, Phase diagram.
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Received 8 November 2016, Accepted 5 January 2017, Published 31 January 2017

© The Japan Society of Microgravity Applicaiton

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