Isolation of indoleamine oxygenase gene from the mollusc Turbo carnutus and functional convergence

• Project/Area Number: 12640664
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 動物生理・代謝
• Research Institution: Kochi University
• Principal Investigator: SUZUKI Tomohiko Kochi Univ., Fac, Sci., Prof., 理学部, 教授 (60145109)
• Project Period (FY): 2000 – 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2002: ¥600,000 (Direct Cost: ¥600,000); Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: myoglobin / hemoglobin / IDO / Turbo / Sulculus / convergent evolution / アルギニンキナーゼ

Research Abstract

Some members of archaeogastropodic molluscs such as Sulculus and Turbo contain unusual 〜40 kDa myoglobin in their buccal masses. The myoglobin can bind oxygen reversibly, although the oxygen affinity is lower than those of vertebrate and invertebrate myoglobins. The amino acid sequencing clearly showed that Sulculus and Turbo myoglobins evolved not from globin gene but from gene for indoleamine dioxygenase (IDO), a tryptophan-degrading enzyme. The structure of Turbo myoglobin gene has been determined to consist of 14 exons and 13 introns. Compared with the known gene for Sulculus IDO-like myoglobin, all splice junctions but one are conserved exactly between two genes. The exon/intron organization of these myoglobin genes is also highly homologous with that (10 exon-9 intron structure) of human IDO : splice junctions of 6 introns were exactly conserved between three genes, suggesting that these introns have been conserved for at least 600 million years. The open reading frame of Turbo myoglobin cDNA was cloned into the plasmids pQE, pGEX and pET-44, and expressed in E. coli. No myoglobin was expressed in use of pQE vector, while a large amount of fusion protein was expressed as insoluble (pGEX) or soluble (pET-44) forms. To look for putative IDO gene in Turbo or Sulculus, we reexamined the genomic DNA fragments amplified by PCR in full detail, and found two distinct Sulculus fragments A and B containing intron 2. The fragment A corresponded exactly to the myoglobin gene of Sulculus, containing 576 bp intron. On the other hand, fragment B, containing 239 bp intron, differed significantly from fragment A in nucleotide and translated amino acid sequences. Detailed sequence comparison suggests that fragment B may be derived from putative IDO gene of Sulculus.

Research Products

• [Publications] Suzuki, T., Yokouchi, K., Kawamichi, H., Yamamoto, Y., Uda, K., Yuasa, H.J.: "Comparison of the sequences of Turbo and Sulculus inodeamine dioxygenase-like myoglobin genes"Gene. 308. 89-94 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Suzuki, T., Takao, R.Yamanaka, K., Gotoh, H., Furukohri, T., Takagi, T.: "Evidence of met-form myoglobin from Theliostyla albicilla radular muscle"Int. J. Biochem. Cell Biol.. 35. 1119-1126 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Suzuki, T., Sugimura, N., et al.: "Two-domain arginine kinases from the clams Solen strictus and Corbicula japonica. Exceptional amino acid replacement of the functionally important D^<62> by G"Int. J. Biochem. Cell Biol.. 34. 1221-1229 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Suzuki, T., Yokouchi, K., Kawamichi, H., Yamamoto, Y., Uda, K. and Yuasa, H. J.: "Comparison of the sequences of Turbo and Sulculus inodeamine dioxygenase-like myoglobin genes"Gene. 308. 89-94 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Suzuki, T., Takao, H., Yamanaka, K., Gotoh, H., Furukohri, T. and Takagi, T.: "Evidence of metform myoglobin from Theliostyla albicilla radular muscle"Int. J. Biochem. Cell Biol.. 35. 1119-1126 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Suzuki, T., Sugimura, N., Taniguchi, T., Unemi, Y., Murata, T., Hayashida, M., Yokouchi, K., Uda, K. and Furukohri, T.: "Two-domain arginine kinases from the clams Solen strictus and Corbicula japonica. Exceptional amino acid replacement of the functionally important D^<62> by G"Int. J. Biochem. Cell Biol.. 34. 1221-1229 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Suzuki, T., Yokouchi, K., Kawamichi, H., Yamamoto, Y., Uda, K., Yuasa, H.J.: "Comparison of the sequences of Turbo and Sulculus inodeamine dioxygenase-Like myoglobin genes"Gene. (in press). (2003)
• [Publications] Suzuki, T., Takao, H., Yamanaka, K., Gotoh, H., Furukohri, T., Takagi, T.: "Evidence of met-form myoglobin from Theliostyla albicilla radular muscle"Int. J. Biochem. Cell Biol.. (in press). (2003)
• [Publications] Kitazoe, Kurihara, Narita, Okuhara, tominaga, Suzuki: "A New Theory of Phylogeny Inference Through Constraction of Multidimensional vector Space"Mol. Biol. Evol.. 18. 812-828 (2001)
• [Publications] Green, Gotoh, Suzuki, Zal, Lallier, Toulmond, Vinagradov: "Electrospray Ionization Mass Spectrometric Observation of Large, Noncornlent Globin Subassemblies in the -3600 kDa Hexagonal Bilayer Hemoglobins"J. Mol. Biol.. 309. 563-570 (2001)
• [Publications] Suzuki T. et al.: "Arginine kinase from Nautilus pompilius, a living fossil"J.Biol.Chem.. 275. 23884-23890 (2000)
• [Publications] Suzuki T. et al.: "Gastropod arginine kinases from Cellana grata and Aplysia kurodai."Com.Biochem.Physiol. (Biochem.& Mol.Biol.). 127. 505-512 (2000)