Expression of Desulfovibrio gigas desulforedoxin in Escherichia coli: Purification and characterization of mixed metal isoforms

Christopher Czaja, Robert Litwiller, Andy J. Tomlinson, Stephen Naylor, Pedro Tavares, Jean LeGall, José J. G. Moura, Isabel Moura, Frank Rusnak

Research output: Contribution to journalArticlepeer-review

32 Citations (Scopus)
8 Downloads (Pure)

Abstract

The dsr gene from Desulfovibrio gigas encoding the nonheme iron protein desulforedoxin was cloned using the polymerase chain reaction, expressed in Escherichia coli, and purified to homogeneity. The physical and spectroscopic properties of the recombinant protein resemble those observed for the native protein isolated from D. gigas. These include an α2 tertiary structure, the presence of bound iron, and absorbance maxima at 370 and 506 nm in the UV/visible spectrum due to ligand-to-iron charge transfer bands. Low temperature electron paramagnetic resonance studies confirm the presence of a high-spin ferric ion with g values of 7.7, 5.7, 4.1, and 1.8. Interestingly, E. coli produced two forms of desulforedoxin containing iron. One form was identified as a dimer with the metal-binding sites of both subunits occupied by iron while the second form contained equivalent amounts of iron and zinc and represents a dimer with one subunit occupied by iron and the second with zinc.

Original languageEnglish
Pages (from-to)20273-20277
Number of pages5
JournalJournal of Biological Chemistry
Volume270
Issue number35
DOIs
Publication statusPublished - 1 Sep 1995

Fingerprint

Dive into the research topics of 'Expression of Desulfovibrio gigas desulforedoxin in Escherichia coli: Purification and characterization of mixed metal isoforms'. Together they form a unique fingerprint.

Cite this