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Expression and characterisation of a major c-type cytochrome encoded by gene kustc0563 from Kuenenia s tuttgartiensis as a recombinant protein in Escherichia coli

Huston, Wilhelmina M., Harhangi, Harry R., Leech, Andrew P., Bulter, Clive S., Jetten, Mike S., Op den Camp, Huub J., & Moir, James W. (2007) Expression and characterisation of a major c-type cytochrome encoded by gene kustc0563 from Kuenenia s tuttgartiensis as a recombinant protein in Escherichia coli. Protein Expression and Purification, 51(1), pp. 28-33.

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Abstract

The purification of small quantities of a major small c-type cytochrome from the anammox bacterium Kuenenia stuttgartiensis has recently been reported. In order to characterise this protein further we have expressed the gene encoding this cytochrome in Escherichia coli and have purified the protein to homogeneity. The protein is directed to the E. coli periplasm using its native signal sequence suggesting that it may be translocated via a Sec-type system in K. stuttgartiensis. The cytochrome has the visible spectroscopic properties typical of a low-spin c-type cytochrome, but these spectroscopic features broaden in high salt solutions. The oxidised cytochrome was able to bind the ligands NO and cyanide. A redox potential of +230 mV suggests that the protein is suitable to act as an electron carrier protein that may be involved in the respiratory chain between hydrazine oxidation and the reduction of nitrite. The predicted protein sequence for the cytochrome suggests it to be a predominantly a-helical protein, and this is supported by circular dichroism.

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ID Code: 7156
Item Type: Journal Article
Additional Information: For more information, please refer to the journal’s website (see link) or contact the author. Author contact details: w.huston@qut.edu.au
Keywords: anammox, cytochrome c, Kuenenia stuttgartiensis
DOI: 10.1016/j.pep.2006.06.026
ISSN: 1046-5928
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > MICROBIOLOGY (060500) > Microbial Genetics (060503)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > MICROBIOLOGY (060500) > Bacteriology (060501)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > BIOCHEMISTRY AND CELL BIOLOGY (060100) > Biochemistry and Cell Biology not elsewhere classified (060199)
Divisions: Past > QUT Faculties & Divisions > Faculty of Science and Technology
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright 2007 Elsevier
Deposited On: 27 Apr 2007
Last Modified: 29 Feb 2012 23:41

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