Spectroscopic characterization of (57)Fe-enriched cytochrome c.
Investigation of the heme iron dynamics in cytochrome c with Mössbauer spectroscopy and especially nuclear resonance vibrational spectroscopy requires the replacement of the natural abundant heme iron with the (57)Fe isotope. For demetallization, we use a safer and milder ferrous sulfate-hydrochloric acid method in addition to the harsher commonly used hydrofluoric acid-based procedure. The structural integrity of the (57)Fe-reconstituted protein in both oxidation states is confirmed from absorption spectra and a detailed analysis of the rich resonance Raman spectra. These results reinforce the application of metal-substituted heme c proteins as reliable models for the native proteins.
Department of Physics and Center for Interdisciplinary Research on Complex Systems, Northeastern University, Boston, MA 02115, USA. leu@aps.anl.gov
Bogdan M
BM
Leu
Tom H
TH
Ching
Cuong
C
Tran
J Timothy
JT
Sage
D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism; D005296:Ferrous Compounds / Q000737:chemistry; D006418:Heme / Q000737:chemistry; D006851:Hydrochloric Acid / Q000737:chemistry; D007503:Iron Isotopes / Q000737:chemistry; D008958:Models, Molecular; D010084:Oxidation-Reduction; D015204:Spectroscopy, Mossbauer; D013059:Spectrum Analysis, Raman; D014732:Vibration
D005296:Ferrous Compounds; D007503:Iron Isotopes; C020748:ferrous sulfate; D006418:Heme; D045304:Cytochromes c; D006851:Hydrochloric Acid