Preparation, Characterization and Stability of Neuroglobin Mutant Tyr44Phe

Abstract

Abstract:

Neuroglobin(Ngb) is a recently discovered member of the hemoglobin superfamily in nervous system of vertebrates. Tyr44 nearing bis-histidyl ligand His64 contacts with one of the heme propionates by hydrogen bonds, which may involue in the stabilization for the hexacoordinate state of neuroglobin. In order to investigate the role of Tyr44 in structural stabilization of neuroglobin, the neuroglobin mutant Tyr44Phe gene was constructed by site-directed mutagenesis, and the mutant protein was expressed, purified and characterized spectroscopicaly, and its stability was also studied. The electrospray ionization mass spectroscopy result show that the molecular weight of the mutant Tyr44Phe corresponds with the theoretical value. The UV-Visible spectra, fluorescence spectra and circular dichroism(CD) spectra of mutant Tyr44Phe indicated that the mutagenesis doesn′t change neuroglobin’s bis-histidyl heme hexacoordination, but has an effect on heme configuration. Results of stability towards heat and acid show that the mutagenesis decreases the interaction of heme with peptide chain and the heme’s hydrophobic cavity becomes flexible, which proves that Tyr44 plays an important role in neuroglobin structural stability.

Key words: Neuroglobin, Mutant Tyr44Phe, Stability

TrendMD:

LI Hai-Li, LI Lian-Zhi*, GUO Yu-Jing, TIAN Su-Yan, XUE Ze-Chun, JIANG Yu-Gang. Preparation, Characterization and Stability of Neuroglobin Mutant Tyr44Phe[J]. Chem. J. Chinese Universities, 2009, 30(5): 928.

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