The Distal Cavity Structure of Carbonyl Horseradish Peroxidase As Probed by the Resonance Raman Spectra of His 42 Leu and Arg 38 Leu Mutants

Feis, Alessandro; Rodríguez-López, José Neptuno; Thorneley, R. N. F.; Smulevich, Giulietta

doi:10.1021/bi981399v

Cited by 42 publications

(63 citation statements)

References 31 publications

(74 reference statements)

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“…The observed ν(Fe-C)/ν(CO) frequencies are typical of haeme-CO adducts that have little interaction with the distal protein matrix. In horseradish peroxidase C (HRP-C), where a distal histidine and a distal arginine interact with the bound CO, the ν(Fe-C) and ν(CO) stretching mode frequencies are found at 539 and 1906 cm − 1 respectively [48]. These findings are in agreement with recent computational studies demonstrating that the mobile distal side Arg 173 points away from the haeme in ferrous NdCld [26].…”

Section: Co Binding To Wild-type Ndcld and The Variants E210a And K141esupporting

confidence: 86%

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

et al. 2016

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SynopsisChlorite dismutase (Cld) and HemQ are structurally and phylogenetically closely related haeme enzymes differing fundamentally in their enzymatic properties. Clds are able to convert chlorite into chloride and dioxygen, whereas HemQ is proposed to be involved in the haeme b synthesis of Gram-positive bacteria. A striking difference between these protein families concerns the proximal haeme cavity architecture. The pronounced H-bonding network in Cld, which includes the proximal ligand histidine and fully conserved glutamate and lysine residues, is missing in HemQ. In order to understand the functional consequences of this clearly evident difference, specific hydrogen bonds in Cld from 'Candidatus Nitrospira defluvii' (NdCld) were disrupted by mutagenesis. The resulting variants (E210A and K141E) were analysed by a broad set of spectroscopic (UV-vis, EPR and resonance Raman), calorimetric and kinetic methods. It is demonstrated that the haeme cavity architecture in these protein families is very susceptible to modification at the proximal site. The observed consequences of such structural variations include a significant decrease in thermal stability and also affinity between haeme b and the protein, a partial collapse of the distal cavity accompanied by an increased percentage of low-spin state for the E210A variant, lowered enzymatic activity concomitant with higher susceptibility to self-inactivation. The high-spin (HS) ligand fluoride is shown to exhibit a stabilizing effect and partially restore wild-type Cld structure and function. The data are discussed with respect to known structure-function relationships of Clds and the proposed function of HemQ as a coprohaeme decarboxylase in the last step of haeme biosynthesis in Firmicutes and Actinobacteria.

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Section: Co Binding To Wild-type Ndcld and The Variants E210a And K141esupporting

confidence: 86%

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

et al. 2016

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“…8 (31). Similar effects are observed for peroxidases (30,32). The distal pockets of peroxidases are typically more polar than that in Mb; the data points therefore fall on the upper left corner of the imidazole correlation curve.…”

Section: ϫ)supporting

confidence: 69%

The Heme Environment of Recombinant Human Indoleamine 2,3-Dioxygenase

Terentis

Thomas

Takikawa

et al. 2002

Journal of Biological Chemistry

144

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“…The electronic absorption and RR spectra of the Fe(II)-CO complex of Ca 2ϩ -depleted HRPC at pH 7.0 (data not shown) show close similarities with the corresponding spectra of the native protein. This indicates that, as in the native protein, two conformers are formed in the CO complex of Ca 2ϩ -depleted HRPC corresponding to hydrogen bonding between CO and either the distal Arg or distal His residue (31).…”

Section: Carbon Monoxide and Bha Binding To Camentioning

confidence: 78%

The Critical Role of the Proximal Calcium Ion in the Structural Properties of Horseradish Peroxidase

Howes

Feis

Raimondi³

et al. 2001

Journal of Biological Chemistry

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The extent to which the structural Ca 2؉ ions of horseradish peroxidase (HRPC) are a determinant in defining the heme pocket architecture is investigated by electronic absorption and resonance Raman spectroscopy upon removal of one Ca 2؉ ion. The Fe(III) heme states are modified upon Ca 2؉ depletion, with an uncommon quantum mechanically mixed spin state becoming the dominant species. Ca 2؉ -depleted HRPC forms complexes with benzohydroxamic acid and CO which display spectra very similar to those of native HRPC, indicating that any changes to the distal cavity structural properties upon Ca 2؉ depletion are easily reversed. Contrary to the native protein, the Ca 2؉ -depleted ferrous form displays a low-spin bis-histidyl heme state and a small proportion of high-spin heme. Furthermore, the (Fe-Im) stretching mode downshifts 27 cm ؊1 upon Ca 2؉ depletion revealing a significant structural perturbation of the proximal cavity near the histidine ligand. The specific activity of the Ca 2؉ -depleted enzyme is 50% that of the native form. The effects on enzyme activity and spectral features observed upon Ca 2؉ depletion are reversible upon reconstitution. Evaluation of the present and previous data firmly favors the proximal Ca 2؉ ion as that which is lost upon Ca 2؉ depletion and which likely plays the more critical role in regulating the heme pocket structural and catalytic properties.Peroxidases of the plant peroxidase superfamily are hemecontaining enzymes that oxidize a variety of aromatic molecules in the presence of hydrogen peroxide. They include peroxidases of plant, fungal, and prokaryotic origins that can be divided into three classes based on sequence alignment (1).Additional support for such a classification was gained as the crystal structures of representative enzymes of each class became available. Class I contains bacterial peroxidases and peroxidases from plant mitochondria and chloroplasts, for example most ascorbate peroxidases and cytochrome c peroxidase. Class II contains extracellular fungal peroxidases such as Coprinus cinereus peroxidase (a peroxidase essentially identical to Arthromyces ramosus peroxidase) and lignin-degrading peroxidases. Class III contains secretory plant peroxidases, typified by the classical horseradish peroxidase isoenzyme C (HRPC). The peroxidases of classes II and III share a number of structural elements considered to be of importance for maintaining protein stability and activity. These include calciumbinding sites proximal and distal to the heme and four disulfide bridges (1-5); the latter was in different locations in class II with respect to class III peroxidases. These features are absent in class I peroxidases. Class III peroxidases also have a loop insertion in the sequence, composed of the DЈ, FЈ, and FЉ helices, not found in the other classes. The relationship between calcium binding and enzyme inactivation has been treated primarily by studies on lignin peroxidase (LIP) (6, 7), manganese peroxidase (MNP) (8, 9), and HRPC (10, 11). Thermal (6) and alkaline (7) i...

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The Distal Cavity Structure of Carbonyl Horseradish Peroxidase As Probed by the Resonance Raman Spectra of His 42 Leu and Arg 38 Leu Mutants

Cited by 42 publications

References 31 publications

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

The Heme Environment of Recombinant Human Indoleamine 2,3-Dioxygenase

The Critical Role of the Proximal Calcium Ion in the Structural Properties of Horseradish Peroxidase

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