title: Cloning, expression and purification of Atlantic salmon (Salmo salar, L.) neuroglobin.
abstract: Neuroglobin (Ngb) exists only in small amounts in salmon brain. In order to study the protein in more detail salmon neuroglobin (sNgb) was cloned, heterologously expressed in Escherichia coli and purified. The protein had red color and showed the characteristic peaks at 411nm (metNgb), 415nm (carboxyNgb) and 424nm (deoxyNgb). Western analysis showed that sNgb reacted weakly against a rabbit anti human neuroglobin (hNgb) and strongly to a sNgb specific antibody. Our 3D-homology model of the sNgb indicated modifications adjacent to and in the O(2)/CO binding site. This may correlate to differences in substrate affinities for the sNgb compared to the hNgb. Also sNgb contained shorter helixes and longer interhelical loops typical for psychrophilic proteins.
authors.affiliation: University of Bergen, Department of Molecular Biology, Bergen, Norway.
authors.forename: Gry Aletta
authors.initials: GA
authors.lastname: Bjørlykke
authors.forename: Bjørn Olav
authors.initials: BO
authors.lastname: Kvamme
authors.forename: Erik
authors.initials: E
authors.lastname: Slinde
authors.forename: Arnt J
authors.initials: AJ
authors.lastname: Raae
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005914:Globins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000078787:Neuroglobin; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020317:Salmo salar; D016415:Sequence Alignment; D013057:Spectrum Analysis; D040681:Structural Homology, Protein
chemical_list: D029941:Fish Proteins; D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D011994:Recombinant Proteins; D005914:Globins