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Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR
ABSTRACT: There is growing experimental evidence that many proteins exhibit a tendency for (ultra)weak homo- or hetero- oligomerization interactions. With the development of paramagnetic relaxation enhancement NMR spectroscopy it has become possible to characterize weak complexes experimentally and...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4908164/ https://www.ncbi.nlm.nih.gov/pubmed/27236778 http://dx.doi.org/10.1007/s10858-016-0035-z |
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author | Schilder, Jesika Ubbink, Marcellus |
author_facet | Schilder, Jesika Ubbink, Marcellus |
author_sort | Schilder, Jesika |
collection | PubMed |
description | ABSTRACT: There is growing experimental evidence that many proteins exhibit a tendency for (ultra)weak homo- or hetero- oligomerization interactions. With the development of paramagnetic relaxation enhancement NMR spectroscopy it has become possible to characterize weak complexes experimentally and even detect complexes with affinities in the 1–25 mM range. We present evidence for a weak complex between cytochrome c peroxidase (CcP) molecules. In a previous study, we attached nitroxide based spin labels at three positions on CcP with the intent of observing intramolecular PRE effects. However, several intermolecular PRE effects were also observed suggesting a weak self-association between CcP molecules. The CcP–CcP complex was characterized using paramagnetic NMR and protein docking. The interaction occurs between the surface that is also part of the stereo-specific binding site for its physiological partner, cytochrome c (Cc), and several small, positively charged patches on the “back” of CcP. The CcP–CcP complex is not a stereo-specific complex. It is a dynamic ensemble of orientations, characteristic of an encounter state. The contact areas resemble those observed for CcP molecules in crystals. The CcP–CcP complex formation competes with that of the CcP-Cc complex. However, the affinity for Cc is much larger and thus it is expected that, under physiological conditions, auto-inhibition will be limited. GRAPHICAL ABSTRACT: A weak self-association between cytochrome c peroxidase molecules was characterized using paramagnetic NMR. [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10858-016-0035-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4908164 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-49081642016-06-30 Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR Schilder, Jesika Ubbink, Marcellus J Biomol NMR Article ABSTRACT: There is growing experimental evidence that many proteins exhibit a tendency for (ultra)weak homo- or hetero- oligomerization interactions. With the development of paramagnetic relaxation enhancement NMR spectroscopy it has become possible to characterize weak complexes experimentally and even detect complexes with affinities in the 1–25 mM range. We present evidence for a weak complex between cytochrome c peroxidase (CcP) molecules. In a previous study, we attached nitroxide based spin labels at three positions on CcP with the intent of observing intramolecular PRE effects. However, several intermolecular PRE effects were also observed suggesting a weak self-association between CcP molecules. The CcP–CcP complex was characterized using paramagnetic NMR and protein docking. The interaction occurs between the surface that is also part of the stereo-specific binding site for its physiological partner, cytochrome c (Cc), and several small, positively charged patches on the “back” of CcP. The CcP–CcP complex is not a stereo-specific complex. It is a dynamic ensemble of orientations, characteristic of an encounter state. The contact areas resemble those observed for CcP molecules in crystals. The CcP–CcP complex formation competes with that of the CcP-Cc complex. However, the affinity for Cc is much larger and thus it is expected that, under physiological conditions, auto-inhibition will be limited. GRAPHICAL ABSTRACT: A weak self-association between cytochrome c peroxidase molecules was characterized using paramagnetic NMR. [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10858-016-0035-z) contains supplementary material, which is available to authorized users. Springer Netherlands 2016-05-28 2016 /pmc/articles/PMC4908164/ /pubmed/27236778 http://dx.doi.org/10.1007/s10858-016-0035-z Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Article Schilder, Jesika Ubbink, Marcellus Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR |
title | Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR |
title_full | Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR |
title_fullStr | Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR |
title_full_unstemmed | Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR |
title_short | Weak self-association of cytochrome c peroxidase molecules observed by paramagnetic NMR |
title_sort | weak self-association of cytochrome c peroxidase molecules observed by paramagnetic nmr |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4908164/ https://www.ncbi.nlm.nih.gov/pubmed/27236778 http://dx.doi.org/10.1007/s10858-016-0035-z |
work_keys_str_mv | AT schilderjesika weakselfassociationofcytochromecperoxidasemoleculesobservedbyparamagneticnmr AT ubbinkmarcellus weakselfassociationofcytochromecperoxidasemoleculesobservedbyparamagneticnmr |