The cytochrome c domain of dimeric cytochrome cd(1) of Paracoccus pantotrophus can be produced at high levels as a monomeric holoprotein using an improved c-type cytochrome expression system in Escherichia coli.
Cytochrome cd(1) nitrite reductase from Paracoccus pantotrophus is a dimer; within each monomer there is a largely alpha-helical domain that contains the c-type cytochrome centre. The structure of this domain changes significantly upon reduction of the heme iron, for which the ligands change from His17/His69 to Met106/His69. Overproduction, using an improved Escherichia coli expression system, of this c-type cytochrome domain as an independent monomer is reported here. The properties of the independent domain are compared with those when it is part of dimeric holo or semi-apo cytochrome cd(1).
Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, United Kingdom.
E H
EH
Gordon
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Steensma
S J
SJ
Ferguson
D003001:Cloning, Molecular; D003574:Cytochrome c Group; D003580:Cytochromes / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D009572:Nitrite Reductases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D010230:Paracoccus / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003574:Cytochrome c Group; D003580:Cytochromes; D011994:Recombinant Proteins; C019315:cytochrome cd1; D009572:Nitrite Reductases