Photo-CIDNP NMR spectroscopy of a heme-containing protein

Day, Iain J.; Wain, Rachel; Tozawa, Keiichi; Smith, Lorna J.; Hore, P. J.

doi:10.1016/j.jmr.2005.04.004

Cited by 5 publications

(2 citation statements)

References 45 publications

(66 reference statements)

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“…In particular, we have used NMR spectroscopy to compare the structure and dynamics (Palmer 2004 ) of the wild-type and b -type variants of the protein. The assignment of the NMR spectrum and some characterisation of the C10A/C13A HT- c 552 have been reported previously (Day et al 2005 ; Pertinhez et al 2001 ; Wain et al 2001 , 2004 ). Here, we have analysed backbone dynamics of WT HT- c 552 and C10A/C13A HT- c 552 and hydrogen exchange protection, along with chemical shift and coupling constant data for these variants, to try to gain insights into the role of the covalently attached heme group in c -type cytochromes.…”

Section: Introductionsupporting

confidence: 60%

Comparison of the backbone dynamics of wild-type Hydrogenobacter thermophilus cytochrome c 552 and its b-type variant

et al. 2015

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Cytochrome c552 from the thermophilic bacterium Hydrogenobacter thermophilus is a typical c-type cytochrome which binds heme covalently via two thioether bonds between the two heme vinyl groups and two cysteine thiol groups in a CXXCH sequence motif. This protein was converted to a b-type cytochrome by substitution of the two cysteine residues by alanines (Tomlinson and Ferguson in Proc Natl Acad Sci USA 97:5156–5160, 2000a). To probe the significance of the covalent attachment of the heme in the c-type protein, 15N relaxation and hydrogen exchange studies have been performed for the wild-type and b-type proteins. The two variants share very similar backbone dynamic properties, both proteins showing high 15N order parameters in the four main helices, with reduced values in an exposed loop region (residues 18–21), and at the C-terminal residue Lys80. Some subtle changes in chemical shift and hydrogen exchange protection are seen between the wild-type and b-type variant proteins, not only for residues at and neighbouring the mutation sites, but also for some residues in the heme binding pocket. Overall, the results suggest that the main role of the covalent linkages between the heme group and the protein chain must be to increase the stability of the protein.

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Section: Introductionsupporting

confidence: 60%

Comparison of the backbone dynamics of wild-type Hydrogenobacter thermophilus cytochrome c 552 and its b-type variant

et al. 2015

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“…Chemically Induced Dynamic Nuclear Polarization (CIDNP) spectroscopy has been widely used as a valuable tool to distinct reaction pathways involving short‐lived radicals, including studies on proteins in terms of mapping of amino‐acid residues towards their accessibility for electron transfer reactions . It has been reported that CIDNP intensities of geminate radical pairs can be translated into hyperfine coupling constants of the two reacting radicals .…”

Section: Introductionmentioning

confidence: 99%

CIDNP as a tool for rapid rearrangements: New insights into cyclobutanoid 4‐center/3‐electron radical cations

Neshchadin

Gescheidt

2013

J of Physical Organic Chem

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CIDNP has been often utilized as a mechanistic tool for radical reactions. Here, we show that detailed insights into the structure of carbon-centered radical cations with unusual structures, which, otherwise, are hardly detectable, can be obtained. Hydrocarbon cage compounds containing syn-periplanar cyclobutane moieties were oxidized by photoinduced one-electron transfer to chloranil at room temperature in CD 2 Cl 2 . The 1 H CIDNP polarization intensities of geminate radical pairs can be translated into isotropic hyperfine coupling constants and reveal radical cations with remarkable electronic structures. Density functional theory calculations are in line with the experimental results. a "Extended"/"Tight", the data for the tight isomer were determined by fluorescence-detected magnetic resonance [19] . b Only protons clearly distinguishable by different chemical shifts are given. c The number in parenthesis gives the average value for b protons resembling the CIDNP polarization intensities (see text). CIDNP OF CAGE COMPOUNDS

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Photo-CIDNP NMR Spectroscopy of Amino Acids and Proteins

Kuhn

2013

Hyperpolarization Methods in NMR Spectroscopy

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Photo-chemically induced dynamic nuclear polarization (CIDNP) is a nuclear magnetic resonance (NMR) phenomenon which, among other things, is exploited to extract information on biomolecular structure via probing solvent-accessibilities of tryptophan (Trp), tyrosine (Tyr), and histidine (His) amino acid side chains both in polypeptides and proteins in solution. The effect, normally triggered by a (laser) light-induced photochemical reaction in situ, yields both positive and/or negative signal enhancements in the resulting NMR spectra which reflect the solvent exposure of these residues both in equilibrium and during structural transformations in "real time". As such, the method can offer - qualitatively and, to a certain extent, quantitatively - residue-specific structural and kinetic information on both the native and, in particular, the non-native states of proteins which, often, is not readily available from more routine NMR techniques. In this review, basic experimental procedures of the photo-CIDNP technique as applied to amino acids and proteins are discussed, recent improvements to the method highlighted, and future perspectives presented. First, the basic principles of the phenomenon based on the theory of the radical pair mechanism (RPM) are outlined. Second, a description of standard photo-CIDNP applications is given and it is shown how the effect can be exploited to extract residue-specific structural information on the conformational space sampled by unfolded or partially folded proteins on their "path" to the natively folded form. Last, recent methodological advances in the field are highlighted, modern applications of photo-CIDNP in the context of biological NMR evaluated, and an outlook into future perspectives of the method is given.

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Photo-CIDNP NMR spectroscopy of a heme-containing protein

Cited by 5 publications

References 45 publications

Comparison of the backbone dynamics of wild-type Hydrogenobacter thermophilus cytochrome c 552 and its b-type variant

Comparison of the backbone dynamics of wild-type Hydrogenobacter thermophilus cytochrome c 552 and its b-type variant

CIDNP as a tool for rapid rearrangements: New insights into cyclobutanoid 4‐center/3‐electron radical cations

Photo-CIDNP NMR Spectroscopy of Amino Acids and Proteins

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