PMID: 1279Jan 2, 1976

Hydrogen-isotope exchange of oxidized and reduced cytochrome c. A comparison of mass spectrometry and infrared methods

European Journal of Biochemistry
E Nabedryk-VialaJ M Thiery

Abstract

Hydrogen-deuterium exchange in 2H20 solutions of the two redox states of horse heart cytochrome c was investigated at 20 degrees C, pH 7, by mass spectrometry and infrared spectroscopy. Mass spectrometry indicates that ferricytochrome has 20 hydrogens unexchanged after 24 h, 28 hydrogens exchanging between 10 min and 24 h, and 156 hydrogens exchanging within 10 min; comparative values for ferrocytochrome are 45, 19 and 140. The displacement of the exchange curves obtained by infrared corresponds to 8 to 9 peptide hydrogens. These combined methods show many non-peptide hydrogens exchanging rapidly (87 and 79 for ferricytochrome c and ferrocytochrome c respectively), whereas others, probably buried inside the molecule and involved in hydrogen bonds, are not exchanged, even after 24 h (14 and 30 hydrogens respectively, which is relatively large for a small protein). Infrared results are given in terms of changes of standard free energy for the transconformational reaction which exposes the peptide hydrogens to solvent: in ferricytochrome c and ferrycoytochrome c, 30% and 40% respectively of the peptide hydrogens are protected by conformational transitions stabilized by more than 5 kcal/mol (21 kJ/mol), which implies a large increa...Continue Reading

References

Feb 18, 1974·Annals of the New York Academy of Sciences·R E Dickerson
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Citations

Nov 25, 1998·Protein Science : a Publication of the Protein Society·L Wang, N R Kallenbach
Jan 1, 1982·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·L Pershina
Jan 1, 1982·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·B A Helm
May 23, 2003·Protein Science : a Publication of the Protein Society·Yong-Sung KimJohn F Carpenter
Oct 30, 1978·Biochemical and Biophysical Research Communications·Y Combarnous, E Nabedryk-Viala
Apr 14, 1980·Biochemical and Biophysical Research Communications·C ThieryJ M Thiery
Feb 4, 2021·European Biophysics Journal : EBJ·Joëlle De Meutter, E Goormaghtigh

Related Concepts

Cytochrome c Group
Hydrogen-Ion Concentration
Mathematics
Myocardium
Oxidation-Reduction
Plasma Protein Binding Capacity
Protein Conformation
Infrared Spectrophotometry
Mass Spectrometry
Thermodynamics

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