Ligand pathways in neuroglobin revealed by low-temperature photodissociation and docking experiments

IUCrJ

卷:6

Thierry Prangé¹  Beatrice Vallone²  Maurizio Brunori²  Linda Celeste Montemiglio²  Giovanna Avella²  Cécile Exertier²  Chiara Ardiccioni³  Stefano Della Longa⁴  Philippe Carpentier⁵  Peter van der Linden⁵  Nathalie Colloc'h⁶  Carmelinda Savino⁷  Alessandro Arcovito⁸  Dominique Bourgeois⁹  Martin Weik⁹ 

[1] CiTeCoM UMR 8038 CNRS, Université Paris Descartes, Paris, France;

[2] Department of Biochemical Sciences `A. Rossi Fanelli', University of Rome Sapienza, Piazzale Aldo Moro 5, 00185 Rome, Italy;

[3] Department of Life and Environmental Sciences, New York–Marche Structural Biology Center (NY-MaSBiC), Polytechnic University of Marche, Ancona, Italy;

[4] Department of Life, Health and Environmental Sciences, University of L'Aquila, 67100 L'Aquila, Italy;

[5] European Synchrotron Radiation Facility (ESRF), 38043 Grenoble, France;

[6] ISTCT UMR 6030 CNRS Université de Caen Normandie CEA, CERVOxy Team, Centre Cyceron, Caen, France;

[7] Institute of Molecular Biology and Pathology, National Research Council, Piazzale Aldo Moro 5, 00185 Rome, Italy;

[8] Istituto di Biochimica e Biochimica Clinica, Universitá Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy;

[9] Université Grenoble Alpes, CEA, CNRS, IBS, 38000 Grenoble, France;

关键词: ultralow-temperature X-ray crystallography;    XANES;    heme protein;    neuroglobin;    oxygen binding;    cryo-trapping;    crystal microspectroscopy;    neuroprotection;    structural biology;    CO photolysis;    soak-and-freeze pressurization;    structure determination;    protein structure;   

DOI  :  10.1107/S2052252519008157

来源: DOAJ

【 摘 要 】

A combined biophysical approach was applied to map gas-docking sites within murine neuroglobin (Ngb), revealing snapshots of events that might govern activity and dynamics in this unique hexacoordinate globin, which is most likely to be involved in gas-sensing in the central nervous system and for which a precise mechanism of action remains to be elucidated. The application of UV–visible microspectroscopy in crystallo, solution X-ray absorption near-edge spectroscopy and X-ray diffraction experiments at 15–40 K provided the structural characterization of an Ngb photolytic intermediate by cryo-trapping and allowed direct observation of the relocation of carbon monoxide within the distal heme pocket after photodissociation. Moreover, X-ray diffraction at 100 K under a high pressure of dioxygen, a physiological ligand of Ngb, unravelled the existence of a storage site for O2 in Ngb which coincides with Xe-III, a previously described docking site for xenon or krypton. Notably, no other secondary sites were observed under our experimental conditions.

【 授权许可】

Unknown