The cDNA-Derived Amino Acid Sequence of Hemoglobin II from Lucina pectinata
Tóm tắt
Hemoglobin II from the clam Lucina pectinata is an oxygen-reactive protein with a unique structural organization in the heme pocket involving residues Gln65 (E7), Tyr30 (B10), Phe44 (CD1), and Phe69 (E11). We employed the reverse transcriptase-polymerase chain reaction (RT-PCR) and methods to synthesize various cDNAHbII. An initial 300-bp cDNA clone was amplified from total RNA by RT-PCR using degenerate oligonucleotides. Gene-specific primers derived from the HbII-partial cDNA sequence were used to obtain the 5′ and 3′ ends of the cDNA by RACE. The length of the HbII cDNA, estimated from overlapping clones, was approximately 2114 bases. Northern blot analysis revealed that the mRNA size of HbII agrees with the estimated size using cDNA data. The coding region of the full-length HbII cDNA codes for 151 amino acids. The calculated molecular weight of HbII, including the heme group and acetylated N-terminal residue, is 17,654.07 Da.
Tài liệu tham khảo
Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W., et al. (1997). Nucleic Acids Res. 25: 3389–3402.
Antommattei-Pérez, F. M., Rosado-Ruiz, T., Cadilla, C. L., and López-Garriga, J. (1999). J. Protein Chem. 18: 831–836.
Brunori, M., Cutruzzola, F., Savino, C., Travaglini-Allocatelli, C., Vallone, B., and Gibson, Q. H. (1999). Trends Biochem. Sci. 24: 223–255.
Chomczynski, P., and Sacchi, N. (1987). Anal. Biochem. 162: 156–159.
Dewilde, S., Winnepenninckx, B., Arndt, M. H. L., Nascimento, D. G., Santoro, M., Knight, M., et al. (1998). J. Biol. Chem. 273: 13583–13592.
Draghi, F., Miele, A. E., Travaglini-Allocatelli, C., Vallone, B., Brunori, M., Gibson, Q. H., et al. (2002). J. Biol. Chem. 277: 7509–7519.
Hockenhull-Johnson, J. D., Stern, M. S., Martin, P., Dass, C., Desiderio, D. M., Wittenberg, J. B., et al. (1991). J. Protein Chem. 10: 609–622.
Hockenhull-Johnson, J. D., Stern, M. S., Wittenberg, J. B., Vinogradov, S. N., Kapp, O. H., and Walz, D. A. (1993). J. Protein Chem. 12: 261–277.
Huang, S., Huang, J., Kloek, A. P., Goldberg, D. E., and Friedman, J. M. (1996). J Biol. Chem. 271: 958–962.
Kraus, D. W., and Wittenberg, J. B. (1990). J. Biol. Chem. 265: 16043–16053.
Kraus, D. W., Wittenberg, J. B., Jing-Fen, L., and Peisach, J. (1990). J. Biol. Chem. 265: 16054–16059.
Mukai, M., Mills, C. E., Poole, R. K., and Yeh, S. (2001). J. Biol. Chem. 276: 7272–7277.
Parente, A., Verde, C., Malorni, A., Montecucchi, P., Aniello, F., and Geraci, G. (1993). Biochim. Biophys. Acta 1162(1–2): 1–9.
Peterson, E. S., Huang, S., Wang, J., Miller, L. M., Vidugiris, G., Kloek, A., et al. (1997). Biochemistry 36: 13110–13121.
Read, K. R. H. (1965). Comp. Biochem. Physiol. 15: 137–158.
Rizzi, M., Wittenberg, J. B., Coda, A., Fasano, M., Ascenzi, P., and Bolognesi, M. (1994). J. Mol. Biol. 244: 86–99.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989). Molecular Cloning—A Laboratory Manual, 2nd Edition. Cold Spring Harbor, New York, Cold Spring Harbor Laboratory Press.
Sanoguet, Z. (1999). M.S. Thesis, University of Puerto Rico, Mayagüez, P.R.
Spencer, N. (1984). J. Mol. Biol. 107: 405–415.
Suzuki, T., Arita, T., and Kawasaki, Y. (1995). Zool. Sci. 12: 453–455.
Takano, T. 1977. J. Mol. Biol. 110: 537–568.