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

IUCrJ

Carpentier, P.¹  Savino, C.²  Ardiccioni, C.³  van der Linden, P.⁴  Bourgeois, D.⁵  Della Longa, S.⁶  Avella, G.⁷  Exertier, C.⁸  Arcovito, A.⁹  Weik, M.^1,10  Montemiglio, L.C.^1,11 

[1] Chemistry Department, Merck Serono S.p.A., Via Casilina 125, 00176 Rome, Italy;Department of Biochemical Sciences `A. Rossi Fanelli', University of Rome Sapienza, Piazzale Aldo Moro 5, 00185 Rome, Italy;Department of Life and Environmental Sciences, New YorkMarche Structural Biology Center (NY-MaSBiC), Polytechnic University of Marche, Ancona, Italy;Department of Life, Health and Environmental Sciences, University of L'Aquila, 67100 L'Aquila, Italy;European Synchrotron Radiation Facility (ESRF), 38043 Grenoble, France;Fondazione Policlinico Universitario Agostino GemelliIRCCS, Largo Francesco Vito 1, 00168 Rome, Italy;Institute of Molecular Biology and Pathology, National Research Council, Piazzale Aldo Moro 5, 00185 Rome, Italy;Istituto PasteurFondazione Cenci Bolognetti, Department of Biochemical Sciences `A. Rossi Fanelli', University of Rome Sapienza, Piazzale Aldo Moro 5, 00185 Rome, Italy;Istituto di Biochimica e Biochimica Clinica, Universit Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy;Partnership for Soft Condensed Matter (PSCM), 38043 Grenoble, France;Universit 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 ST;   

DOI  :  10.1107/S2052252519008157

学科分类：数学（综合）

来源: International Union of Crystallography

【 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】

CC BY