Clarification of hemoglobin evolution process by reverse molecular evolution

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This study intends to learn the principle of protein molecule design which was adopted by nature to acquire high-order functions for ancestral hemoglobins of vertebrates in the course of molecular evolution. We designed and synthesized the hemoglobin genes for ancestors of teleost and cyclostomata and expressed them in E. coli, obtaining the ancestral hemoglobins. The teleost ancestral hemoglobins showed a strong Bohr effect which assumes a Root effect and week cooperativity, indicating that they have characteristics of teleost. The cyclostomata ancestral hemoglobins showed a small Bohr effect and week cooperativity in contrast with natural lamprey hemoglobin without those effects.