A proton NMR study of the non-covalent complex of horse cytochrome c and yeast cytochrome-c peroxidase and its comparison with other interacting protein complexes.
Abstract: Cytochrome-c peroxidase (ferrocytochrome-c:hydrogen-peroxide oxidoreductase, EC 1.11.1.5) forms a noncovalent 1:1 complex with horse cytochrome c in low ionic strength solution that is detectable by proton NMR spectroscopy. When the entire proton hyperfine-shifted spectrum is considered only five hyperfine resonances exhibit unambiguously detectable shifts: the heme 8-CH3 and 3-CH3 resonances, single proton resonances near 19 ppm and -4 ppm and the methionine-80 methyl group. These shifts are very similar to those observed for the covalently crosslinked complex of cytochrome-c peroxidase and horse cytochrome c, but different from those reported for cytochrome c complexes with flavodoxin and cytochrome b5. By comparison with the shifts reported for lysine-13-modified cytochrome c we conclude that the results reported here support the Poulos-Kraut proposed structure for the molecular redox complex between cytochrome-c peroxidase and cytochrome c. These results indicate that the principal site of interaction with cytochrome-c peroxidase is the exposed heme edge of horse cytochrome c, in proximity to lysine-13 and the heme pyrrole II. The noncovalent cytochrome-c peroxidase-cytochrome c complex exists in the rapid-exchange time limit even at 500 mHz proton frequency. Our data provide an improved estimate of the minimum off-rate for exchanging cytochrome c as 1133 (+/- 120) s-1 at 23 degrees C.
Publication Date: 1987-03-18 PubMed ID: 3030433DOI: 10.1016/0167-4838(87)90251-2Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Research Support
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research study uses proton NMR spectroscopy to examine the non-covalent complex formed between horse cytochrome c and yeast cytochrome-c peroxidase. The findings provide evidence supporting a specific proposed structure for the molecule complex and identify the main site of interaction, which helps in understanding the redox complex.
Understanding the Non-Covalent Complex
- The researchers investigated the non-covalent 1:1 complex that forms between horse cytochrome c and yeast cytochrome-c peroxidase in low ionic strength solution and it was detected and examined by proton NMR spectroscopy.
- They noticed that only five hyperfine resonances show unambiguously detectable shifts. These include; the heme 8-CH3 and 3-CH3 resonances, single proton resonances near 19 ppm and -4 ppm, and the methionine-80 methyl group.
- These identified shifts were discovered to be very similar to those seen in the covalently crosslinked complex of cytochrome-c peroxidase and horse cytochrome c, but they were found to be different from those reported for cytochrome c complexes with flavodoxin and cytochrome b5.
Comparison and Conclusions
- The observed shifts aided in supporting the Poulos-Kraut proposed structure for the molecular redox complex existing between cytochrome-c peroxidase and cytochrome c.
- The results indicate that the principal site of interaction with cytochrome-c peroxidase is the exposed heme edge of horse cytochrome c, in proximity to lysine-13 and the heme pyrrole II.
- Moreover, it was noticed that the non-covalent cytochrome-c peroxidase-cytochrome c complex exists in the rapid-exchange time limit even at 500 mHz proton frequency. This conclusion was significant to the study as it provided an improved estimate of the minimum off-rate for exchanging cytochrome c as 1133 (+/- 120) s-1 at 23 degrees C.
Cite This Article
APA
Satterlee JD, Moench SJ, Erman JE.
(1987).
A proton NMR study of the non-covalent complex of horse cytochrome c and yeast cytochrome-c peroxidase and its comparison with other interacting protein complexes.
Biochim Biophys Acta, 912(1), 87-97.
https://doi.org/10.1016/0167-4838(87)90251-2 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Computer Graphics
- Cytochrome c Group
- Cytochrome-c Peroxidase
- Heme
- Horses
- Lysine
- Magnetic Resonance Spectroscopy
- Models, Molecular
- Peroxidases
- Protein Binding
- Protein Conformation
- Saccharomyces cerevisiae / enzymology
Grant Funding
- RR1081 / NCRR NIH HHS
Citations
This article has been cited 7 times.- van Son M, Schilder JT, Di Savino A, Blok A, Ubbink M, Huber M. The Transient Complex of Cytochrome c and Cytochrome c Peroxidase: Insights into the Encounter Complex from Multifrequency EPR and NMR Spectroscopy.. Chemphyschem 2020 May 18;21(10):1060-1069.
- Page TR, Hoffman BM. Control of cyclic photoinitiated electron transfer between cytochrome c peroxidase (W191F) and cytochrome c by formation of dynamic binary and ternary complexes.. Biochemistry 2015 Feb 10;54(5):1188-97.
- Moore GR, Cox MC, Crowe D, Osborne MJ, Rosell FI, Bujons J, Barker PD, Mauk MR, Mauk AG. N epsilon,N epsilon-dimethyl-lysine cytochrome c as an NMR probe for lysine involvement in protein-protein complex formation.. Biochem J 1998 Jun 1;332 ( Pt 2)(Pt 2):439-49.
- Satterlee JD, Moench S. Proton hyperfine resonance assignments using the nuclear Overhauser effect for ferric forms of horse and tuna cytochrome c.. Biophys J 1987 Jul;52(1):101-7.
- Weber C, Michel B, Bosshard HR. Spectroscopic analysis of the cytochrome c oxidase-cytochrome c complex: circular dichroism and magnetic circular dichroism measurements reveal change of cytochrome c heme geometry imposed by complex formation.. Proc Natl Acad Sci U S A 1987 Oct;84(19):6687-91.
- Dixon DW, Hong X, Woehler SE. Electrostatic and steric control of electron self-exchange in cytochromes c, c551, and b5.. Biophys J 1989 Aug;56(2):339-51.
- Busse SC, Moench SJ, Satterlee JD. One- and two-dimensional proton NMR studies of cys-102 S-methylated yeast isozyme-1 ferricytochrome c.. Biophys J 1990 Jul;58(1):45-51.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
