Properties of catalase-peroxidase lacking its C-terminal domain

Biochemical and Biophysical Research Communications
Ruletha D BakerDouglas C Goodwin

Abstract

Catalase-peroxidases have a two-domain structure. The N-terminal domain contains the bifunctional active site, but the function of the C-terminal domain is unknown. We produced catalase-peroxidase containing only its N-terminal domain (KatG(Nterm)). Removal of the C-terminal domain did not result in unexpected changes in secondary structure as evaluated by CD, but KatG(Nterm) had neither catalase nor peroxidase activity. Partial recovery of both activities was achieved by incubating KatG(Nterm) with the separately expressed and isolated KatG C-terminal domain. Spectroscopic measurements revealed a shift in heme environment from a mixture of high-spin species (wtKatG) to exclusively hexacoordinate, low-spin (KatG(Nterm)). Moreover, a > 1000-fold lower kon for CN- binding was observed for KatG(Nterm). EPR spectra for KatG(Nterm) and the results of site-specific substitution of active site histidines suggested that the distal histidine was the sixth ligand. Thus, one important role for the C-terminal domain may be to support the architecture of the active site, preventing heme ligation by this catalytically essential residue.

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Citations

Nov 28, 2013·Archives of Biochemistry and Biophysics·Olive J NjumaDouglas C Goodwin
May 24, 2008·Antioxidants & Redox Signaling·Marcel ZamockyChristian Obinger
Jul 27, 2010·Biochimica Et Biophysica Acta·Srijib BanerjeeChristian Obinger
Oct 7, 2009·Free Radical Biology & Medicine·Tomoya YataYoshinobu Takakura
Jan 8, 2010·Protein Science : a Publication of the Protein Society·Christine E CadeReza A Ghiladi
Apr 17, 2008·Biochimica Et Biophysica Acta·Robert L MooreDouglas C Goodwin
Dec 5, 2006·Biophysical Journal·Inés García-RubioArthur Schweiger
Aug 21, 2020·Nature Communications·Souvik BhattacharyyaRasika M Harshey
Jan 7, 2018·Archives of Biochemistry and Biophysics·Vanessa Vega-GarcíaWilhelm Hansberg
Mar 7, 2006·Journal of Inorganic Biochemistry·Giulietta SmulevichChristian Obinger

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