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Journal of Protein Chemistry

, Volume 15, Issue 8, pp 709–719 | Cite as

Primary structure of mannuronate lyases SP1 and SP2 fromTurbo cornutus and involvement of the hydrophobic C-terminal residues in the protein stability

  • Tsuyoshi Muramatsu
  • Kenji Komori
  • Narumi Sakurai
  • Koji Yamada
  • Yasuyuki Awasaki
  • Kazumasa Fukuda
  • Tatsuya Oda
Article

Abstract

The complete amino acid sequences of two isoforms, SP1 and SP2, of mannuronate lyase from a wreath shell,Turbo cornutus, were determined to elucidate amino acid residues responsible for causing the more stable protein conformation of SP2. The sequences of the two isoforms were identical except for two hydrophobic C-terminal amino acid residues of SP2, Ile and Leu, which were additionally attached to Thr of the C-terminal residue of SP1 (253 residues in total). The molecular weight of SP2 was calculated to be 28,912 from the amino acid sequence data. Two disulfide bond cross-linkages were found to be between 106 and 115 and between 145 and 150, and a partially buried single SH group was located at 236. A carbohydrate chain that consisted of 3 GlcNAc, 3 Fuc, and 1 Man was anchored on Asn-105 in a typical carbohydrate-binding motif of Asn-X-Ser. This is the first evidence of the primary structure of mannuronate lyase, and no significant homology of the amino acid sequence among other proteins was found. The C-terminal truncated SP2, which was produced by digestion with carboxypeptidase Y and corresponded structurally to SP1, showed a thermal stability identical to that of SP1. These results indicate that the higher stability of SP2 than SP1 arises from the presence of the C-terminal two hydrophobic amino acid residues.

Key words

Mannuronate lyase amino acid sequence stability disulfide bond C-terminal residue 

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References

  1. Crestfield, A. M., Moore, S., and Stein, W. H. (1963).J. Biol. Chem. 238, 622–627.PubMedGoogle Scholar
  2. Edelhoch, H. (1967).Biochemistry 6, 1948–1954.PubMedGoogle Scholar
  3. Gacesa, P. (1992).Int. J. Biochem. 24, 545–552.PubMedGoogle Scholar
  4. Kellis, J. T., Nyberg, K., Sali, D., and Fersht, A. R. (1988).Nature 333, 784–786.PubMedGoogle Scholar
  5. Matsumura, M., Yahanda, S., Yasumura, S., Yutani, K., and Aiba, S. (1988).Eur. J. Biochem. 171, 715–720.PubMedGoogle Scholar
  6. Moore, S. (1963).J. Biol. Chem. 238, 235–237.Google Scholar
  7. Muramatsu, T., and Egawa, K. (1982).Agric. Biol. Chem. 46, 883–889.Google Scholar
  8. Muramatsu, T., and Imasato, F. (1987).Agric. Biol. Chem. 51, 1169–1171.Google Scholar
  9. Muramatsu, T., Yamada, K., Date, M., and Yoshioka, S. (1993).Biosci. Biotechnol. Biochem. 57, 1990–1994.Google Scholar
  10. Muramatsu, T., Hashimoto, H., and Takahashi, T. (1984).Agric. Biol. Chem. 48, 79–85.Google Scholar
  11. Van der Schueren, J., Robben, J., Goossens, K., Heremans, K., and Volckaert, G. (1996).J. Mol. Biol. 256, 878–888.PubMedGoogle Scholar
  12. Yutani, K., Ogasahara, K., Tsujita, T., and Sugino, Y. (1987).Proc. Natl. Acad. Sci. USA. 84, 4441–4444.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • Tsuyoshi Muramatsu
    • 1
  • Kenji Komori
    • 1
  • Narumi Sakurai
    • 1
  • Koji Yamada
    • 1
  • Yasuyuki Awasaki
    • 1
  • Kazumasa Fukuda
    • 1
  • Tatsuya Oda
    • 1
  1. 1.Division of Biochemistry, Faculty of FisheriesNagasaki UniversityNagasakiJapan

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