Biophysical Characterisation of Neuroglobin of the Icefish, a Natural Knockout for Hemoglobin and Myoglobin. Comparison with Human Neuroglobin

• Main Authors: Giordano, D., Boron, I., Abbruzzetti, S., van Leuven, W., Nicoletti, F.P., Forti, F., Bruno, S., Cheng, C.-H.C., Moens, L., di Prisco, G., Nadra, A.D., Estrin, D., Smulevich, G., Dewilde, S., Viappiani, C., Verde, C.
• Format: Journal/Newspaper
• Language: unknown
• Subjects: hemoglobin; myoglobin; neuroglobin; article; autooxidation; biophysics; Chaenocephalus aceratus; controlled study; Dissostichus mawsoni; fish; human; human versus animal comparison; laser flash photolysis; molecular cloning; molecular dynamics; nonhuman; oxygen affinity; oxygen transport; photolysis; protein analysis; protein expression; protein function; protein purification; protein structure; Raman spectrometry; site directed mutagenesis; ultraviolet spectroscopy; Animals; Carbon Monoxide; Fishes; Gene Knockout Techniques; Globins; Hemoglobins; Humans; Kinetics; Ligands; Molecular Dynamics Simulation; Mutagenesis; Site-Directed; Nerve Tissue Proteins; Spectrophotometry; Ultraviolet; Spectrum Analysis; Raman; Vertebrata; Antarc*; Antarctic; Icefish
• Online Access: https://hdl.handle.net/20.500.12110/paper_19326203_v7_n12_p_Giordano

The Antarctic icefish Chaenocephalus aceratus lacks the globins common to most vertebrates, hemoglobin and myoglobin, but has retained neuroglobin in the brain. This conserved globin has been cloned, over-expressed and purified. To highlight similarities and differences, the structural features of the neuroglobin of this colourless-blooded fish were compared with those of the well characterised human neuroglobin as well as with the neuroglobin from the retina of the red blooded, hemoglobin and myoglobin-containing, closely related Antarctic notothenioid Dissostichus mawsoni. A detailed structural and functional analysis of the two Antarctic fish neuroglobins was carried out by UV-visible and Resonance Raman spectroscopies, molecular dynamics simulations and laser-flash photolysis. Similar to the human protein, Antarctic fish neuroglobins can reversibly bind oxygen and CO in the Fe 2+ form, and show six-coordination by distal His in the absence of exogenous ligands. A very large and structured internal cavity, with discrete docking sites, was identified in the modelled three-dimensional structures of the Antarctic neuroglobins. Estimate of the free-energy barriers from laser-flash photolysis and Implicit Ligand Sampling showed that the cavities are accessible from the solvent in both proteins. Comparison of structural and functional properties suggests that the two Antarctic fish neuroglobins most likely preserved and possibly improved the function recently proposed for human neuroglobin in ligand multichemistry. Despite subtle differences, the adaptation of Antarctic fish neuroglobins does not seem to parallel the dramatic adaptation of the oxygen carrying globins, hemoglobin and myoglobin, in the same organisms. © 2012 Giordano et al. Fil:Nadra, A.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Estrin, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.