Cloning, sequencing and functional expression in Escherichia coli of dmc gene encoding periplasmic tetraheme cytochrome c3 from Desulphovibrio desulphuricans M6

Si Keun Lim, Doo Hyun Park, Yong Keun Park, Byung Hong Kim

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A small soluble protein, periplasmic tetraheme cytochrome c3, was purified from Desulphovibrio desulfuricans M6 and its gene, dmc, was cloned and the complete nucleotide sequence determined. The purity index of purified cytochrome c3 was 3.3 and the molecular weight was determined as 14.5 kDa by SDS-PAGE. It was found that the 387bp of dmc gene encoded 21 amino acids of hydrophobic signal peptide and 107 residues of apoprotein. The nucleotide sequence and the predicted amino acid sequence of dmc showed 76% and 83% identities to those of 13 kDa cytochrome c3 from D. desulphuricans ATCC 27774, respectively, dmc gene was functionally expressed in aerobically grown Escherichia coli BL-21(DE3) by co-expressing eight ccm genes which were reported to be involved in cytochrome c maturation. The molecular weight of overexpressed holocytochrome c3 was identical to that of the original protein. Visible spectrum of dithionite-reduced form exhibited typical characteristics of c-type cytochromes. In addition, the redox potential was measured to -340mV by cyclic voltammetry.

Original languageEnglish
Pages (from-to)263-269
Number of pages7
JournalAnaerobe
Volume7
Issue number5
DOIs
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Organism Cloning
Escherichia coli
Genes
Molecular Weight
Periplasmic Proteins
Cytochrome c Group
Dithionite
Apoproteins
Protein Sorting Signals
Cytochromes c
Oxidation-Reduction
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Amino Acids
cytochrome c(3)
Proteins

Keywords

  • Desulphovibrio desulphuricans M6
  • dmc gene
  • Functional expression
  • Purification
  • Tetraheme cytochrome c

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Cloning, sequencing and functional expression in Escherichia coli of dmc gene encoding periplasmic tetraheme cytochrome c3 from Desulphovibrio desulphuricans M6. / Lim, Si Keun; Park, Doo Hyun; Park, Yong Keun; Hong Kim, Byung.

In: Anaerobe, Vol. 7, No. 5, 2001, p. 263-269.

Research output: Contribution to journalArticle

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