Metal ion coordination sites in ferrochelatase

Hunter, Gregory A.; Ferreira, Glória C.

doi:10.1016/j.ccr.2022.214464

Cited by 11 publications

(10 citation statements)

References 123 publications

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“…This out‐of‐plane deformation exposes both the protons and the lone electron pairs of the nitrogen atoms of the porphyrin core, facilitating the metal insertion. 50 , 51 , 52 This porphyrin deformation was further investigated by site‐directed mutagenesis in murine ferrochelatase. In fact, the variants with a low catalytic efficiency also exhibited a decrease in the intensity of RR out‐of‐plane vibrational mode γ 15 mode, suggesting that the initial PPIX saddling deformation could be directly correlated with catalytic affinity and in particular with metal selectivity.…”

Section: Discussionmentioning

confidence: 99%

“…Based on the structure of the N ‐MeMP transition state analog bound to Bs CpfC taken as reference model, the preponderance of saddling distortion observed upon binding of the free substrate into the protein, has been considered particularly relevant to initiate the fC mechanism. This out‐of‐plane deformation exposes both the protons and the lone electron pairs of the nitrogen atoms of the porphyrin core, facilitating the metal insertion 50–52 . This porphyrin deformation was further investigated by site‐directed mutagenesis in murine ferrochelatase.…”

Section: Discussionmentioning

confidence: 99%

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Active site architecture of coproporphyrin ferrochelatase with its physiological substrate coproporphyrin III: Propionate interactions and porphyrin core deformation

et al. 2022

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Coproporphyrin ferrochelatases (CpfCs) are enzymes catalyzing the penultimate step in the coproporphyrin-dependent (CPD) heme biosynthesis pathway, which is mainly utilized by monoderm bacteria. Ferrochelatases insert ferrous iron into a porphyrin macrocycle and have been studied for many decades, nevertheless many mechanistic questions remain unanswered to date. Especially CpfCs, which are found in the CPD pathway, are currently in the spotlight of research. This pathway was identified in 2015 and revealed that the correct substrate for these ferrochelatases is coproporphyrin III (cpIII) instead of protoporphyrin IX, as believed prior the discovery of the CPD pathway. The chemistry of cpIII, which has four propionates, differs significantly from protoporphyrin IX, which features two propionate and two vinyl groups.These findings let us to thoroughly describe the physiological cpIIIferrochelatase complex in solution and in the crystal phase. Here, we present the first crystallographic structure of the CpfC from the representative monoderm pathogen Listeria monocytogenes bound to its physiological substrate, cpIII, together with the in-solution data obtained by resonance Raman and UV-vis spectroscopy, for wild-type ferrochelatase and variants, analyzing propionate interactions. The results allow us to evaluate the porphyrin distortion and provide an in-depth characterization of the catalytically-relevant binding mode of cpIII prior to iron insertion. Our findings are discussed in the light of the observed structural restraints and necessities for this porphyrin-enzyme complex to catalyze the iron insertion process. Knowledge about this initial situation is essential for understanding the preconditions for iron insertion in CpfCs and builds the basis for future studies.Andrea Dali, Thomas Gabler, and Federico Sebastiani contributed equally to this study.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Active site architecture of coproporphyrin ferrochelatase with its physiological substrate coproporphyrin III: Propionate interactions and porphyrin core deformation

et al. 2022

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“…Iron porphyrin complexes are often active centers of biological enzymes . In order to eliminate the interference of the apoenzyme and effectively improve the catalytic and ultrasensitive detection performance, studying the relationship between the charge-transfer (CT) process and oxidative stress in vivo using an iron porphyrin as an artificial catalytic acceptor to simulate metalloenzymes is of great significance. , Recently, two-dimensional (2D) metal–organic frameworks (MOFs) have stimulated significant attention for their efficient catalysis due to the great active centers from numerous metal nodes and bridging ligands.…”

Section: Introductionmentioning

confidence: 99%

SERS Tracking Oxidative Stress on a Metalloporphyrin Framework by Vitamin C

Xie,

Liu,

Wen

et al. 2023

Anal. Chem.

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Accurate control of charge transfer is crucial to investigate the catalytic reaction mechanism of the biological oxidation process that biomedicine participates in. Herein, we have established an assembly model of metalloporphyrin framework (MPF) nanosheets as the active centers of biological enzymes. The introduction of Vitamin C (VC) into the MPF system can precisely modulate its content of charges. The surface-enhanced Raman scattering activity and peroxidase-like catalytic performance are enhanced simultaneously for the first time by manipulating the optimal molar ratio of an MPF to VC and the reaction sequence with target model molecules. We have confirmed that the formation of the intermediate of Fe( 2+ )-OOH species is specifically enhanced after VC modulation, which indicates that VC can regulate the oxidative stress of the active center of biological enzymes. This discovery not only accurately resolves the mechanism of VC-selective anticancer therapy but also has important significance for the precise treatment of VC synergistic targeting medicines.

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“…Eukaryotic HemH homologues, denoted FECH, particularly those from Saccharomyces cerevisiae and Homo sapiens, are well-studied. − Ec HemH is a membrane-associated enzyme and has been previously expressed and purified, , but no kinetic characterization of Ec HemH, or HemH from any other Gram-negative bacteria, has been reported. The location of the productive metal binding site and the face of the porphyrin into which the metal is inserted have been debated in the literature. , Nevertheless, in vitro incorporation of non-native metals by several ferrochelatases (HemH homologues) is well known. ,− To date, it is unclear how chelatases tune their activity for different metals, and many potential effects may contribute depending on the specific protein and metal in question. Distortion of the porphyrin, − specificity-determining active site residues, , differential product inhibition, and exogenous metal delivery are suggested to contribute.…”

mentioning

confidence: 99%

“…The location of the productive metal binding site and the face of the porphyrin into which the metal is inserted have been debated in the literature. 25,28 Nevertheless, in vitro incorporation of nonnative metals by several ferrochelatases (HemH homologues) is well known. 25,29−32 To date, it is unclear how chelatases tune their activity for different metals, and many potential effects may contribute depending on the specific protein and metal in question.…”

mentioning

confidence: 99%

Molecular Determinants of Efficient Cobalt-Substituted Hemoprotein Production in E. coli

Weaver,

Perkins,

Fernandez Candelaria

et al. 2023

ACS Synth. Biol.

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Exchanging the native iron of heme for other metals yields artificial metalloproteins with new properties for spectroscopic studies and biocatalysis. Recently, we reported a method for the biosynthesis and incorporation of a non-natural metallocofactor, cobalt protoporphyrin IX (CoPPIX), into hemoproteins using the common laboratory strain Escherichia coli BL21(DE3). This discovery inspired us to explore the determinants of metal specificity for metallocofactor biosynthesis in E. coli. Herein, we report detailed kinetic analysis of the ferrochelatase responsible for metal insertion, EcHemH (E. coli ferrochelatase). This enzyme exhibits a small, less than 2-fold preference for Fe 2+ over the non-native Co 2+ substrate in vitro. To test how mutations impact EcHemH, we used a surrogate metal specificity screen to identify variants with altered metal insertion preferences. This engineering process led to a variant with an ∼30-fold shift in specificity toward Co 2+ . When assayed in vivo, however, the impact of this mutation is small compared to the effects of alteration of the external metal concentrations. These data suggest that incorporation of cobalt into PPIX is enabled by the native promiscuity of EcHemH coupled with BL21's impaired ability to maintain transition-metal homeostasis. With this knowledge, we generated a method for CoPPIX production in rich media, which yields cobalt-substituted hemoproteins with >95% cofactor purity and yields comparable to standard expression protocols for the analogous native hemoproteins.

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Metal ion coordination sites in ferrochelatase

Cited by 11 publications

References 123 publications

Active site architecture of coproporphyrin ferrochelatase with its physiological substrate coproporphyrin III: Propionate interactions and porphyrin core deformation

Active site architecture of coproporphyrin ferrochelatase with its physiological substrate coproporphyrin III: Propionate interactions and porphyrin core deformation

SERS Tracking Oxidative Stress on a Metalloporphyrin Framework by Vitamin C

Molecular Determinants of Efficient Cobalt-Substituted Hemoprotein Production in E. coli

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