The effect of iron to manganese substitution on microperoxidase 8 catalysed peroxidase and cytochrome P450 type of catalysis

Primus, Jean-Louis; Boersma, Marelle G.; Mandon, Dominique; Boeren, Sjef; Veeger, C.; Weiss, Raymond; Rietjens, Ivonne M. C. M.

doi:10.1007/s007750050313

Cited by 40 publications

(57 citation statements)

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“…Peroxidase activity of heme peptides was determined essentially as described by Primus et al (27). A total of 1 mM 3,3Ј-dimethoxybenzidine was used as substrate.…”

Section: Methodsmentioning

confidence: 99%

Biosynthesis of artificial microperoxidases by exploiting the secretion and cytochrome c maturation apparatuses of Escherichia coli

Braun¹,

Thöny‐Meyer²

2004

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

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Microperoxidases were initially isolated as peptide fragments containing covalently bound heme and are derived from naturally occurring c-type cytochromes. They are not only used as model compounds but also have potential applications as biosensors, electron carriers, photoreceptors, microzymes, and drugs. In a systematic attempt to define the minimal requirements for covalent attachment of hemes to c-type cytochromes, we have succeeded to produce artificial microperoxidases with peptide sequences that do not occur naturally and can be manipulated. The in vivo production of these microperoxidases requires targeting of the peptide to the bacterial periplasm, proteolytic processing of the signal peptide, and covalent attachment of heme to the signature motif CXXCH by the cytochrome c maturation proteins CcmA-H. The peptides that bind heme carry a C-terminal histidine tag, presumably to stabilize the heme peptide. We present a heme cassette that is the basis for the de novo design of functional hemoproteins.

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“…Peroxidase activity of heme peptides was determined essentially as described by Primus et al (27). A total of 1 mM 3,3Ј-dimethoxybenzidine was used as substrate.…”

Section: Methodsmentioning

confidence: 99%

Biosynthesis of artificial microperoxidases by exploiting the secretion and cytochrome c maturation apparatuses of Escherichia coli

Braun¹,

Thöny‐Meyer²

2004

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

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“…Spectra were externally calibrated with bovine cytochrome c (12,230.9 Da), bovine insulin (5,734.6 Da) (both Sigma), and Microperoxidase 8 (MP8, 1,506.5 Da; Ref. 30).…”

Section: Methodsmentioning

confidence: 99%

Natural Disulfide Bond-disrupted Mutants of AVR4 of the Tomato Pathogen Cladosporium fulvum Are Sensitive to Proteolysis, Circumvent Cf-4-mediated Resistance, but Retain Their Chitin Binding Ability

Burg

Westerink

Françoijs

et al. 2003

Journal of Biological Chemistry

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111

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The extracellular AVR4 elicitor of the pathogenic fungus Cladosporium fulvum induces defense responses in the tomato genotype Cf-4. Here, the four disulfide bonds of AVR4 were identified as Cys-11-41, Cys-21-27, Cys-35-80, and Cys-57-72 by partial reduction with Tris-(2-carboxyethyl)-phosphine hydrochloride, subsequent cyanylation, and base-catalyzed chain cleavage. The resulting peptide fragments were analyzed by mass spectrometry. Sequence homology and the disulfide bond pattern revealed that AVR4 contains an invertebrate (inv) chitin-binding domain (ChBD). Binding of AVR4 to chitin was confirmed experimentally. The three disulfide bonds encompassing the inv ChBD motif are also required for protein stability of AVR4. Independent disruption of each of the three conserved disulfide bonds in AVR4 resulted in a protease-sensitive protein, whereas the fourth disulfide bond appeared not to be required for protein stability. Most strains of C. fulvum virulent on Cf-4 tomato contain Cys to Tyr substitutions in AVR4 involving two (Cys-11-41, Cys-35-80) of the three disulfide bonds present in the inv ChBD motif. These natural Cys to Tyr mutant AVR4 proteins did retain their chitin binding ability and when bound to chitin were less sensitive to proteases. Thus, the widely applied tomato Cf-4 resistance gene is circumvented by C. fulvum by amino acid substitutions affecting two disulfide bonds in AVR4 resulting in the absence of the corresponding AVR4 isoforms in apoplastic fluid. However, these natural isoforms of AVR4 appear to have retained their intrinsic function, i.e. binding to chitin present in the cell wall of C. fulvum, most likely to protect it against the deleterious effects of plant chitinases.Gene-for-gene-based disease resistance in plants commonly requires two complementary genes, an avirulence (Avr) 1 gene in the pathogen and a matching resistance gene in the host (1, 2). The Cf resistance genes of tomato mediate specific recognition of extracellular elicitor proteins encoded by Avr genes of the pathogenic fungus Cladosporium fulvum (3). The Avrs of C. fulvum and their matching Cf genes have become valuable instruments to investigate signal transduction pathways leading to plant disease resistance (4 -7, 9).2 To obtain sustainable resistance, the Cf resistance genes were introgressed from wild Lycopersicon species into commercial tomato cultivars. However, because of selection pressure new strains of C. fulvum emerged that had overcome the introgressed resistance traits by modification of the Avr genes (10). Although some Avr genes in these virulent C. fulvum strains were found to be absent (11), others contained point mutations (12) or transposon insertions (13). The natural strains of C. fulvum carrying these mutated Avr genes did not exhibit significantly reduced virulence under laboratory conditions (11-14), suggesting that AVR proteins are not essential for virulence or that the modified isoforms of AVRs still contribute to virulence of C. fulvum. The genetic variation is so far strictly limited to t...

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“…The second oxidation equivalent is delocalized as a radical on an organic moiety, generally the porphyrin macrocycle [13,14] or a protein residue [15,16,17]. For cytochromes P450 the involvement of an isoelectronic form of peroxidase Compound I is postulated in most of the reactions [18,19,20], but involvement of a porphyrin iron-hydroperoxo intermediate or a porphyrin iron-peroxo intermediate is also proposed [12,21,22,23,24,25,26,27].…”

Section: Introductionmentioning

confidence: 99%

“…Fe(III)MP-8 consists of an iron protoporphyrin IX connected to an octapeptide via two thioether bonds [10]. The distal heme ligand can be easily exchanged for an oxygen donor such as H 2 O 2 , leading to the formation of reactive intermediates able to act in both peroxidase and cytochrome P450 model reactions ( [1,11,12] and references therein). This makes Fe(III)MP-8 an appealing model for studying peroxidase-and cytochrome P450-type catalysis.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of a microperoxidase-8 with a modified histidine axial ligand

et al. 2002

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Microperoxidase-8, Fe(III)MP-8, the heme octapeptide obtained by horse heart cytochrome c digestion, was studied in the presence of H(2)O(2). A modified form of the catalyst was isolated by HPLC and showed a UV/visible spectrum similar to that of Fe(III)MP-8. ESI-MS measurements revealed a 16 Da increase in molecular mass for the modified catalyst when compared to Fe(III)MP-8, suggesting the insertion of an oxygen atom. ESI-MS(2) fragmentation measurements point at oxygen incorporation on the His18 residue of the octapeptide of the modified catalyst. Comparison of the (1)H NMR chemical shifts of the methyl protons of the porphyrin ring of Fe(III)MP-8 and the modified catalyst shows a large shift for especially the 3-methyl and 5-methyl resonances, whereas the other (1)H NMR chemical shifts are almost unaffected. These observations can best be ascribed to a reorientation of the histidine axial ligand. The latter is suggested to be the consequence of an oxygen insertion, possibly on the imidazole ring of His18, thereby corroborating the data obtained by ESI-MS(2). (1)H NMR NOE difference measurements on Fe(III)MP-8 and on the modified catalyst supported the assignment of the H(delta)2 and H(epsilon)1 protons of the His18 imidazole ring. The ring amine proton H(delta)1 could not be detected in both forms of the catalyst. For Fe(III)MP-8 this absence of the H(delta)1 resonance can be ascribed to fast H/D exchange. For the modified catalyst the NMR data are not contradictory, with an oxygen insertion on position delta1 of the His18 imidazole ring with a fast H/D exchanging hydroxyl proton. Together these data converge in suggesting the H(2)O(2) modified catalyst bears a hydroxylated His18 axial ligand. The mechanism that could underlie Fe(III)MP-8 axial histidine hydroxylation is further discussed.

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The effect of iron to manganese substitution on microperoxidase 8 catalysed peroxidase and cytochrome P450 type of catalysis

Cited by 40 publications

References 0 publications

Biosynthesis of artificial microperoxidases by exploiting the secretion and cytochrome c maturation apparatuses of Escherichia coli

Biosynthesis of artificial microperoxidases by exploiting the secretion and cytochrome c maturation apparatuses of Escherichia coli

Natural Disulfide Bond-disrupted Mutants of AVR4 of the Tomato Pathogen Cladosporium fulvum Are Sensitive to Proteolysis, Circumvent Cf-4-mediated Resistance, but Retain Their Chitin Binding Ability

Isolation and characterization of a microperoxidase-8 with a modified histidine axial ligand

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