Photoreduction and validation of haem-ligand intermediate states in protein crystals by in situ single-crystal spectroscopy and diffraction

IUCrJ
Demet KekilliMichael A Hough

Abstract

Powerful synergies are available from the combination of multiple methods to study proteins in the crystalline form. Spectroscopies which probe the same region of the crystal from which X-ray crystal structures are determined can give insights into redox, ligand and spin states to complement the information gained from the electron-density maps. The correct assignment of crystal structures to the correct protein redox and ligand states is essential to avoid the misinterpretation of structural data. This is a particular concern for haem proteins, which can occupy a wide range of redox states and are exquisitely sensitive to becoming reduced by solvated electrons generated from interactions of X-rays with water molecules in the crystal. Here, single-crystal spectroscopic fingerprinting has been applied to investigate the laser photoreduction of ferric haem in cytochrome c'. Furthermore, in situ X-ray-driven generation of haem intermediates in crystals of the dye-decolourizing-type peroxidase A (DtpA) from Streptomyces lividans is described.

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Citations

Jul 12, 2017·Dalton Transactions : an International Journal of Inorganic Chemistry·Amanda K ChaplinJonathan A R Worrall
Mar 2, 2019·Acta Crystallographica. Section D, Structural Biology·Ali EbrahimRobin L Owen
Apr 5, 2019·Chemistry : a European Journal·Amanda K ChaplinJonathan A R Worrall
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Nov 5, 2019·Biochimica Et Biophysica Acta. General Subjects·Michihiro SugaHiroshi Sugimoto
Sep 25, 2020·Archives of Biochemistry and Biophysics·Poonam DhankharPravindra Kumar
Oct 14, 2017·Chemical Reviews·Damián Alvarez-PaggiDaniel H Murgida
Aug 30, 2021·Current Opinion in Structural Biology·Michael A Hough, Robin L Owen
Sep 4, 2021·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Marina LučićJonathan A R Worrall

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Methods Mentioned

BETA
X-ray
single-crystal resonance

Software Mentioned

XDS
AIMLESS
CCP
SpectraSuite
Coot
MOLREP
OriginPro
APE
SpeCuBE
SLS

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