Journal of Protein Chemistry

, Volume 15, Issue 8, pp 755–761 | Cite as

cDNA sequence analysis and mutagenesis studies on the a chain of Β-bungarotoxin from Taiwan banded krait

  • Long-Sen Chang
  • Pei-Fung Wu
  • Chun-Chang Chang
Article

Abstract

The cDNA encoding the A chain ofΒ-bungarotoxin (Β-Bgt) was constructed from the cellular RNA isolated from the venom glands ofBungarus multicinctus (Taiwan banded krait). The deduced amino acid sequence encoding the A chain revealed that the determined chain was different from the known A chains (A1, A2, A3, A4, and A5). Nevertheless, the amino acid sequence and the cDNA sequence of the novel A chain were highly homologous with those of other A chains. The gene encoding the A chain ofΒ-Bgt was subjected to mutagenesis, and the Tyr-11, Cys-15, and Leu-72 of the A chain were substituted by Cys-11, Ser-15, and Cys-72, respectively. Instead of the six disulfide bonds observed with the A chain, the resulting mutant contained seven disulfide linkages in its molecular structure which simulated those of presynaptic PLA2 neurotoxins and PLA2 enzymes. However, the mutant did not exhibit a higher phospholipase activity than that noted with the recombinant A chain. These results seem to suggest that, in the absence of the B chain, the six pairs of disulfide bonds in the recombinant A-chain molecule are enough to maintain its active conformation for exerting the phospholipase activity.

Key words

A chain ofΒ-bungarotoxin cDNA sequence analysis mutagenesis of cysteine residues refolding of the recombinant A chain 

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Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • Long-Sen Chang
    • 1
  • Pei-Fung Wu
    • 1
  • Chun-Chang Chang
    • 1
  1. 1.Department of BiochemistryKaohsiung Medical CollegeKaohsiungTaiwan 807, ROC

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