Proton NMR comparison of noncovalent and covalently cross-linked complexes of cytochrome c peroxidase with horse, tuna, and yeast ferricytochromes c

Moench, Susan J.; Chroni, Stamatia; Lou, Bih Show; Erman, James E.; Satterlee, James D.

doi:10.1021/bi00129a015

Cited by 49 publications

(65 citation statements)

References 71 publications

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“…From these calculations, we conclude that the proteins spend Ͼ70% of the lifetime of the complex in the dominant orientation. This finding is consistent with other studies that have proposed the existence of a single-orientation complex in solution (10,34,35). In particular, large NMR chemical-shift perturbations of Cc resonances upon complex formation suggest the presence of a dominant orientation (30) because for more dynamic protein complexes, the perturbations tend to be smaller (36,37).…”

Section: Materials and Methods)supporting

confidence: 92%

“…The presence of multiple orientations within Cc-CcP complex previously has been suggested by several studies (9)(10)(11)(12)(13)(14)(15)17). A complex comprising an equilibrium between a well defined form and a dynamic state (Fig.…”

Section: Resultsmentioning

confidence: 80%

“…However, it has remained a matter of debate whether this structure represents the only form in solution (6)(7)(8). According to several studies, the complex is dynamic, and the crystal structure might represent only a subpopulation of protein orientations (9)(10)(11)(12)(13)(14)(15). Recent studies show that the photoinduced ET between Znsubstituted CcP and Cc, both in the crystal (8,16) and in solution (17), occurs with faster backward than forward rates, indicating that the complex is present in multiple forms, only a few of which are ET-active (17).…”

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confidence: 99%

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Solution structure and dynamics of the complex between cytochromecand cytochromecperoxidase determined by paramagnetic NMR

Volkov

Worrall

Holtzmann

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

244

361

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The physiological complex of yeast cytochrome c peroxidase and iso-1-cytochrome c is a paradigm for biological electron transfer. Using paramagnetic NMR spectroscopy, we have determined the conformation of the protein complex in solution, which is shown to be very similar to that observed in the crystal structure [ electron transfer ͉ encounter state ͉ transient complex ͉ spin label ͉ paramagnetic relaxation enhancement T he process of protein complex formation can be described by a two-step model, in which a short-lived, dynamic encounter complex precedes a dominant, well defined state (Fig. 1). The former enables proteins to undergo reduced-dimensionality search of the optimal binding geometry, thereby accelerating molecular association as compared with 3D diffusion (1). Fast molecular association is essential for protein-protein complexes that require high turnover rates, like those involved in electron transfer (ET) in photosynthesis, respiration, and other metabolic processes (2). The physiological complex of yeast iso-1-cytochrome c (Cc) and yeast cytochrome c peroxidase (CcP) is a paradigm for the intermolecular ET (3) and is one of the few transient ET complexes for which a crystal structure has been solved (4). A recent study has confirmed that the protein-protein orientation observed in the crystal is ET-active (5). However, it has remained a matter of debate whether this structure represents the only form in solution (6-8). According to several studies, the complex is dynamic, and the crystal structure might represent only a subpopulation of protein orientations (9-15). Recent studies show that the photoinduced ET between Znsubstituted CcP and Cc, both in the crystal (8, 16) and in solution (17), occurs with faster backward than forward rates, indicating that the complex is present in multiple forms, only a few of which are .Characterization of the binding interface in the dynamic encounter state (Fig. 1B) has so far proven to be elusive. Investigation of protein complexes by using x-ray crystallography or conventional NMR spectroscopy addresses only the singleorientation species (Fig. 1C), and the only way to visualize the dynamic state is offered by theoretical modeling studies (11,18). In the recent, elegant work of Clore and coworkers (19,20), it was shown how paramagnetic relaxation can be applied to study the dynamic state of protein-DNA complexes. We report on the application of an analogous experimental approach that allows us to define both the dominant protein-protein orientation (Fig. 1C) and the conformational space sampled by the proteins in the dynamic encounter complex (Fig. 1B). Results and DiscussionSolution Structure of the Complex. The concept of the approach is that NMR resonance intensities of one of the proteins in the complex are affected by a paramagnetic spin label covalently attached to the other protein (Fig. 2). The paramagnetic effects are converted into distance restraints (21-23), which can be used to calculate protein-protein orientations within the complex (24). Five ...

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Section: Materials and Methods)supporting

confidence: 92%

Section: Resultsmentioning

confidence: 80%

mentioning

confidence: 99%

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Solution structure and dynamics of the complex between cytochromecand cytochromecperoxidase determined by paramagnetic NMR

Volkov

Worrall

Holtzmann

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

244

361

View full text Add to dashboard Cite

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“…The resonance of heme methyl 3 is strongly downfield-shifted in yeast cytochrome c when it binds to the yeast peroxidase, whereas that of heme methyl 8 experiences a small upfield shift (27,28). Perturbations of the heme methyl resonances of horse cytochrome c are much smaller (29) and indeed can be mimicked by binding of polyglutamate (30).…”

Section: Discussionmentioning

confidence: 99%

The Structure of an Electron Transfer Complex Containing a Cytochrome c and a Peroxidase

Pettigrew

Prazeres

Costa

et al. 1999

Journal of Biological Chemistry

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“…Early NMR spectroscopy studies have demonstrated that the dissociation rate of the complex is concentration dependent (Moench et al, 1992;Yi et al, 1994a), indicating that complex formation is not a simple bimolecular process. The first two-dimensional mapping of the binding site on Cc was performed by taking advantage of the protection against solvent exchange of amide protons in the interface upon complex formation.…”

Section: A Model For Complex Formationmentioning

confidence: 99%

Transient complexes of redox proteins: structural and dynamic details from NMR studies

Prudêncio

Ubbink

2004

J of Molecular Recognition

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Redox proteins participate in many metabolic routes, in particular those related to energy conversion. Protein-protein complexes of redox proteins are characterized by a weak affinity and a short lifetime. Two-dimensional NMR spectroscopy has been applied to many redox protein complexes, providing a wealth of information about the process of complex formation, the nature of the interface and the dynamic properties of the complex. These studies have shown that some complexes are non-specific and exist as a dynamic ensemble of orientations while in other complexes the proteins assume a single orientation. The binding interface in these complexes consists of a small hydrophobic patch for specificity, surrounded by polar, uncharged residues that may enhance dissociation, and, in most complexes, a ring or patch of charged residues that enhances the association by electrostatic interactions. The entry and exit port of the electrons is located within the hydrophobic interaction site, ensuring rapid electron transfer from one redox centre to the next.

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Proton NMR comparison of noncovalent and covalently cross-linked complexes of cytochrome c peroxidase with horse, tuna, and yeast ferricytochromes c

Cited by 49 publications

References 71 publications

Solution structure and dynamics of the complex between cytochromecand cytochromecperoxidase determined by paramagnetic NMR

Solution structure and dynamics of the complex between cytochromecand cytochromecperoxidase determined by paramagnetic NMR

The Structure of an Electron Transfer Complex Containing a Cytochrome c and a Peroxidase

Transient complexes of redox proteins: structural and dynamic details from NMR studies

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