Expression and mutagenesis studies of cobrotoxin from Taiwan cobra

Long Sen Chang, Ku Chung Chen, Bin Nan Wu, Shu Kai Lin, Pei Fung Wu, Yi Ren Hong, Chen Chung Yang

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The cDNA encoding cobrotoxin was constructed from the cellular RNA isolated from the venom glands of Naja naja atra (Taiwan cobra). The cDNA was subcloned into the expression vector pET20b(+)) and transformed into BL21(DE3) Escherichia coli strain. Expressed cobrotoxin was isolated from inclusion bodies of E. coli and subjected to refolding into its folded structure. The refolded cobrotoxin was purified by high-performance liquid chromatography and exhibited a neurotoxicity in inhibiting acetylcholine-induced muscle contractions. Recombinant cobrotoxin showed a tendency to isomerize its disulfide bonds as that observed with native cobrotoxin. An appreciable decrease in the rate of isomerization reaction was observed when Glu-38 was replaced with Gln-38 or Lys-47 was replaced with Glu-47 or Gln-47. These results reflect that the element in controlling the disulfide isomerization of cobrotoxin is closely associated with the charged side chains in the cobrotoxin molecule.

Original languageEnglish
Pages (from-to)652-656
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume263
Issue number3
DOIs
Publication statusPublished - Oct 5 1999
Externally publishedYes

Fingerprint

Cobra Neurotoxin Proteins
Elapidae
Mutagenesis
Taiwan
Isomerization
Disulfides
Escherichia coli
Complementary DNA
Inclusion Bodies
Venoms
High performance liquid chromatography
Muscle Contraction
Acetylcholine
Muscle
High Pressure Liquid Chromatography
RNA
Molecules

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Expression and mutagenesis studies of cobrotoxin from Taiwan cobra. / Chang, Long Sen; Chen, Ku Chung; Wu, Bin Nan; Lin, Shu Kai; Wu, Pei Fung; Hong, Yi Ren; Yang, Chen Chung.

In: Biochemical and Biophysical Research Communications, Vol. 263, No. 3, 05.10.1999, p. 652-656.

Research output: Contribution to journalArticle

Chang, Long Sen ; Chen, Ku Chung ; Wu, Bin Nan ; Lin, Shu Kai ; Wu, Pei Fung ; Hong, Yi Ren ; Yang, Chen Chung. / Expression and mutagenesis studies of cobrotoxin from Taiwan cobra. In: Biochemical and Biophysical Research Communications. 1999 ; Vol. 263, No. 3. pp. 652-656.
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AU - Hong, Yi Ren

AU - Yang, Chen Chung

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