Abstract
A detailed electrospray ionization mass spectrometric study of the ∼3.5-MDa hexagonal bilayer hemoglobin (HBL Hb) from the pond leech Macrobdella decora has shown it to consist of at least six ∼17-kDa globin chains, of which two are monomeric and the remaining four occur as disulfide-bonded heterodimers, and three ∼24-kDa nonglobin linker chains (Weber et al., J. Mol. Biol. 251: 703–720, 1995). The cDNA sequences of the five major constituent chains, globin chains IIA, IIB, B, and C and linker chain L1, are reported here. The globins and linkers share 30%–50% and 20%–30% identity, respectively, with other annelid sequences. Furthermore, IIB and C align with strain A of annelid sequences, whereas IIA and B align with the strain B sequences. Although chains B and C are monomeric, chains IIA and IIB form the main disulfide-bonded dimer. They also have some unusual features: the distal His (E7) is replaced by Phe in IIA, and the highly conserved CD1Phe is replaced by Leu in IIB. In spite of these unusual features, the functional properties of Macrobdella Hb are comparable to those of other HBL Hbs. A phylogenetic analysis of the globin sequences from Macrobdella, the polychaete Tylorrhynchus, the oligochaete Lumbricus, and the vestimentiferan Lamellibrachia, indicates that the two strains originated by gene duplication followed by additional duplication of each of the two strains. The mutation rate of the linkers appeared to be faster than that of the globin chains. The phylogenetic trees constructed using the Maximum Likelihood, Neighbor-Joining and Fitch methods showed the Macrobdella globin sequences to be closest to Lumbricus, in agreement with a view of annelid evolution in which the divergence of the polychaetes occurred before the divergence of the leeches from oligochaetes.
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REFERENCES
Bashford, D., Chothia, C., and Lesk, A. M. (1987). J. Mol. Biol. 196: 199–216.
Brinkhurst, R. O. (1994). Megadrilogica 5: 109–116.
De Haas, F., Boisset, N., Taveau, J. C., Lambert, O., Vinogradov, S. N., and Lamy, J. N. (1996). Biophys. J. 70: 1973–1984.
Dewilde, S., Van Hauwaert, M. L., Vinogradov, S. N., Vanfleteren, J., and Moens, L. (2001). Biochem. Biophys. Res. Commun. 281: 18–24.
Eibye-Jacobsen, D., and Nielsen, C. (1997). Zool. Scr. 25: 275–282.
Felsenstein, J. (1993) Distributed by the author. Department of Genetics, University of Washington, Seattle.
Ferraguti, M., and Erseus, C. (1999). Hydrobiologia 402: 225–237.
Fushitani, K., Matsuura, M. S., and Riggs, A. F. (1988). J. Biol. Chem. 263: 6502–6517.
Goodman, M., Weiss, M. L., and Czelusniak, J. (1982). Syst. Zool. 31: 376–399.
Gotoh, T., and Suzuki, T. (1990). Zool. Sci. 7: 1–16.
Gotoh, T., Shishikura, F., Snow, J. W., Ereifej, K. I., Vinogradov, S. N., and Walz, D. A. (1987). Biochem. J. 241: 441–445.
Gotoh, T., Sano, T., Shibuya, A., Yamaki, M., Imai, K., and Ebina, S. (1998). Arch. Biochem. Biophys. 360: 75–84.
Green, B. N., Suzuki, T., Gotoh, T., Kuchumov, A. R., and Vinogradov, S. N. (1995). J. Biol. Chem. 270: 18209–18211.
Green, B. N., Kuchumov, A. R., Walz, D. A., and Vinogradov, S. N. (1998). Biochemistry 37: 6598–6605.
Green, B. N., Kuchumov, A. R., Klemm, D., and Vinogradov, S. N. (1999a). Int. J. Mass Spectrosc. Ion Proc. 188: 105–112.
Green, B. N., Bordoli, R. S., Hanin, L., Lallier, F. H., Toulmond, A., and Vinogradov, S. N. (1999b). J. Biol. Chem. 274: 28206–28212.
Green, B. N., Zal, F., Lallier, F. H., Toulmond, A., Suzuki, T., Gotoh, T., et al. (2001). J. Mol. Biol. 309: 553–560.
Kapp, O. H., Qabar, A. N., Bonner, M. C., Stern, M. S., Walz, D. A., Schmuck, M., et al. (1990). J. Mol. Biol. 213: 141–158.
Kapp, O. H., Moens, L., Vanfleteren, J., Trotman, C. N. A., Suzuki, T., and Vinogradov, S. N. (1995). Protein Sci. 4: 2179–2190.
Kojima, S. (1998). Mol. Phylogenet. Evol. 9: 255–261.
Kuchumov, A. R., Taveau, J.-C., Lamy, J. N., Wall, J. S., Weber, R. E., and Vinogradov, S. N. (1999). J. Mol. Biol. 289: 1361–1374.
Lamy, J., Kuchumov, A. R., Taveau, J.-C., Boisset, N., Vinogradov, S. N., and Lamy, J. N. (2000). J. Mol. Biol. 298: 633–647.
Lamy, J. N., Green, B. N., Toulmond, A., Wall, J. S., Weber, R. E., and Vinogradov, S. N. (1996). Chem. Rev. 96: 3113–3124.
Maier, C. S., Arbogast, B., Hahn, A., Kuchumov, A. R., Vinogradov, S. N., Walz, D. A., et al. (1997). J. Am. Soc. Mass Spectrom. 21: 563–574.
Martin, P., Kuchumov, A. R., Braswell, E., Green, B. N., Oliver, R. M. W., Wall, J. S., et al. (1996). J. Mol. Biol. 255: 154–169.
McHugh, D. (1997). Proc. Natl. Acad. Sci. USA 94: 8006–8009.
Moens, L., Vanfleteren, J., Van de Peer, Y., Peeters, K., Kapp, O. H., Czeluzniak, J., et al. (1996). Mol. Biol. Evol. 13: 324–333.
Negrisolo, E., Pallavicini, A., Barbato, R., Dewilde, S., Ghiretti-Magaldi, A., Moens, L., et al. (2001). J. Biol. Chem. 276: 26391–26397.
Pallavicini, A., Negrisolo, E., Barbaot, R., Dewilde, S., Ghiretti-Magaldi, A., Moens, L., et al. (2001). J. Biol. Chem. 276: 26384–26390.
Pesce, A., Couture, M., Dewilde, S., Guertin, M., Yamauchi, K., Ascenzi, P., et al. (2000). EMBO J. 11: 2424–2434.
Phillips, S. E. V., and Schoenborn, B. P. (1981). Nature 292: 81–82.
Purschke, G., Westheide, W., Rohde, D., and Brinkhurst, R. O. (1993). Zoomorphology 113: 91–101.
Qabar, A. N., Stern, M. S., Walz, D. A., Chiu, J. T., Timkovich, R. S., Wall, J. S., et al. (1991). J. Mol. Biol. 222: 1109–1129.
Rouse, G. W., and Fauchald, K. (1995). Zool. Scr. 24: 269–301.
Rouse, G. W., and Fauchald, K. (1997). Zool. Scr. 26: 139–204.
Royer, W. E., Jr., Strand, K., van Heel, M., and Hendrickson, W. A. (2000). Proc. Natl. Acad. Sci. USA 97: 7107–7111.
Shishikura, F., and Nakamura, A. (1996). Zool. Sci. 13: 849–856.
Shishikura, F., Snow, J. S., Gotoh, T., Vinogradov, S. N., and Walz, D. A. (1987). J. Biol. Chem. 262: 3123–3131.
Shishikura, F., Ochiai, T., and Yamanaka, I. (1997). Zool. Sci 14: 923–930.
Shlom, J. M., and Vinogradov, S. N. (1973). J. Biol. Chem. 248: 7904–7912.
Siddall, M. E., and Burreson, E. M. (1995). Can. J. Zool. 73: 1048–1064.
Siddall, M. E., and Burreson, E. M. (1996). Hydrobiologia 334: 277–285.
Siddall, M. E., and Burreson, E. M. (1998). Mol. Phylogenet. Evol. 9: 156–162.
Simons, P. C., and Satterlee, J. D. (1989). Biochemistry 28: 8525–8530.
Stern, M. S., Vinogradov, S. N., Sharma, P. K., Ereifej, K., and Walz, D. A. (1990). Eur. J. Biochem. 194: 67–73.
Strimmer, K., and Von Haeseler, A. (1997) PUZZLE version 4.0. Distributed by the author. Zoologisches Institut, Universität Muenchen, Muenchen, Germany.
Sudhof, T. C., Goldstein, J. L., Brown, M. S., and Russell, D. W. (1985). Science 228: 815–822.
Suzuki, T. (1989). Eur. J. Biochem. 185: 127–134.
Suzuki, T., and Gotoh, T. (1986). J. Biol. Chem. 261: 9257–9267.
Suzuki, T., and Riggs, A. F. (1993). J. Biol. Chem. 268: 13548–13555.
Suzuki, T., Kapp, O. H., and Gotoh, T. (1988). J. Biol. Chem. 263: 18524–18529.
Suzuki, T., Takagi, T., and Ohta, S. (1990a). Biochem. J. 26: 221–225.
Suzuki, T., Takagi, T., and Gotoh, T. (1990b). J. Biol. Chem. 265: 12168–12177.
Suzuki, T., Takagi, T., and Ohta, S. (1990c). J. Biol. Chem. 265: 1551–1555.
Suzuki, T., Ohta, T., Yuasa, J., and Takagi, T. (1994). Biochim. Biophys. Acta 1217: 291–296.
Suzuki, T., Hirao, Y., and Vinogradov, S. N. (1995). Biochim. Biophys. Acta 1252: 189–193.
Takagi, T., Iwaasa, H., Ohta, S., and Suzuki, T. (1991). In Vinogradov, S. N., and Kapp, O. H. (eds.), Structure and Function of Invertebrate Oxygen Carriers, Springer, New York, pp. 245–249.
Vinogradov, S. N. (1985a). Comp. Biochem. Physiol. 82B: 1–15.
Vinogradov, S. N. (1985b). In Lamy, J., Truchot, J. P., and Gilles, R. (eds.), Respiratory Pigments in Animals, Springer, Berlin, pp. 9–20.
Vinogradov, S. N., Kapp, O. H., and Ohtsuki, M. (1982). In Harris, J. (ed.), Electron Microscopy of Proteins, Vol. 3, Academic Press, New York, pp. 135–163.
Vinogradov, S. N., Standley, P. R., Mainwaring, M. G., Kapp, O. H., and Crewe, A. V. (1985). Biochim. Biophys. Acta 828: 43–50.
Vinogradov, S. N., Sharma, P. K., and Walz, D. A. (1991a). Comp. Biochem. Physiol. 98B: 187–194.
Vinogradov, S. N., Sharma, P. K., Qabar, A. N., Wall, J. S., Westrick, J. A., Simmons, J. H., et al. (1991b). J. Biol. Chem. 266: 13091–13096.
Vinogradov, S. N., Walz, D. A., Pohajdak, B., Moens, L., Kapp, O. H., Suzuki, T., et al. (1993). Comp. Biochem. Physiol. 106B: 1–26.
Weber, R. E., and Vinogradov, S. N. (2001). Physiol. Rev. 81: 568–629.
Weber, R. E., Malte, H., Sharma, P. K., Green, B. N., Oliver, R. M. W., Braswell, E., et al. (1995). J. Mol. Biol. 251: 703–720.
Westheide, W. (1997). J. Nat. Hist. 31: 1–15.
Westheide, W., McHugh, D., Purschke, G., and Rouse, G. (1999). Hydrobiologia 402: 291–307.
Wittenberg, J. B., Bolognesi, M., Wittenberg, B. A., and Guertin, M. (2002). J. Biol. Chem. 277: 871–874.
Xie, Q., Donahue, R. A., Jr., Schneider, K., Mirza, U. A., Haller, I., Chait, B. T., et al. (1997). Biochim. Biophys. Acta 1337: 241–247.
Yuasa, H. J., Takagi, T., Suzuki, N., Vinogradov, S. N., and Suzuki, T. (1996). Biochim. Biophys. Acta 1296: 235–244.
Zal, F., Lallier, F. H., Wall, J. S., Vinogradov, S. N., and Toulmond, A. (1996a). J. Biol. Chem. 271: 8869–8874.
Zal, F., Lallier, F. H., Green, B. N., Vinogradov, S. N., and Toulmond, A. (1996b). J. Biol. Chem. 271: 8875–8881.
Zal, F., Suzuki, T., Kawasaki, Y., Childress, J. J., Lallier, F. H., and Toulmond, A. (1997). Proteins: Struct. Funct. Genet. 29: 562–574.
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Suzuki, T., Vinogradov, S.N. Globin and Linker Sequences of the Giant Extracellular Hemoglobin from the Leech Macrobdella decora. J Protein Chem 22, 231–242 (2003). https://doi.org/10.1023/A:1025064318790
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DOI: https://doi.org/10.1023/A:1025064318790