Preparation and characterization of a chimeric zebrafish-human neuroglobin engineered by module substitution.
Neuroglobin (Ngb) is a recently discovered vertebrate heme protein that can reversibly bind oxygen that is expressed in the brain. Zebrafish and human Ngb share about 50% amino acid sequence identity. These Ngb proteins consist of four compact protein structural unit "modules" referred to as M1-M4. In the present study, we investigated the effects of module substitution on the properties of Ngb. Specifically, we prepared and characterized a chimeric ZHZZ Ngb in which the heme-binding module M2 of zebrafish Ngb was replaced by the comparable human Ngb module. Our results showed that the chimeric ZHZZ was stable and formed almost the identical heme-environmental and alpha-helical structure as the human and zebrafish Ngb proteins, suggesting that the structure of Ngb has been evolutionarily conserved.
Wakasugi
Keisuke
K
Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan. kei@wakasugi.mbox.media.kyoto-u.ac.jp
Morishima
Isao
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002942:Circular Dichroism; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005914:Globins / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D000078787:Neuroglobin; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D015027:Zebrafish
D017931:DNA Primers; D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D011993:Recombinant Fusion Proteins; D005914:Globins