Spectroscopic identification of the heme axial ligation of cytochrome <i>b</i><sub>558</sub> in the NADPH oxidase of porcine neutrophils

doi:10.1016/0014-5793(95)01372-5

Cited by 14 publications

(2 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Upon reduction with dithionite, the electronic spectra of the wild-type and F190I mutant proteins at pH 5.0 display a prominent absorption band at 557 nm with shoulders near 515 and 585 nm (Figure ), characteristic of pentacoordinate ferrous heme proteins (). When the pH is raised to pH 7.5 for F190I and pH 9.0 for wild-type MnP, the spectra display two prominent absorption bands at 558 and 529 nm, similar to the optical spectrum of ferrous cytochrome b 558 (Table ) (), a known bis-His coordinated iron heme protein ( , ), strongly suggesting the formation of a bis-His heme iron at high pH.…”

Section: Discussionmentioning

confidence: 67%

“…Similar LS bis-His species are formed in both the wild-type and F190I mutant MnP proteins, although the pH at which this transition occurs is significantly lower in the F190I MnP variant. The optical spectra of the reduced proteins at high pH closely resemble those of ferrous cytochrome b 558 (), a known bis-His-ligated protein ( , ). Moreover, the RR spectra of the oxidized and reduced MnP proteins are also similar to those of cytochrome b 558 and bis(imidazole) heme ( 42, 43, 59 ).…”

Section: Discussionmentioning

confidence: 85%

See 1 more Smart Citation

Formation of a Bis(histidyl) Heme Iron Complex in Manganese Peroxidase at High pH and Restoration of the Native Enzyme Structure by Calcium

et al. 2000

View full text Add to dashboard Cite

Manganese peroxidase (MnP) from Phanerochaete chrysosporium undergoes a pH-dependent conformational change evidenced by changes in the electronic absorption spectrum. This high-to lowspin alkaline transition occurs at ∼2 pH units lower in an F190I mutant MnP when compared to the wild-type enzyme. Herein, we provide evidence that these spectral changes are attributable to the formation of a bis(histidyl) heme iron complex in both proteins at high pH. The resonance Raman (RR) spectra of both ferric proteins at high pH are similar, indicating similar heme environments in both proteins, and resemble that of ferric cytochrome b 558 , a protein that contains a bis-His iron complex. Upon reduction with dithionite at high pH, the visible spectra of both the wild-type and F190I MnP exhibit absorption maxima at 429, 529, and 558 nm, resembling the absorption spectrum of ferrous cytochrome b 558 . RR spectra of the reduced wild-type and F190I mutant proteins at high pH are also similar to the RR spectrum of ferrous cytochrome b 558 , further suggesting that the alkaline low-spin species is a bis(histidyl) heme derivative. No shift in the low-frequency RR bands was observed in 75% 18 O-labeled water, indicating that the low-spin species is most likely not a hydroxo-heme derivative. Electronic and RR spectra also indicate that addition of Ca 2+ to either the ferric or ferrous enzymes at high pH completely restores the high-spin pentacoordinate species. Other divalent metals, such as Mn 2+ , Mg 2+ , Zn 2+ , or Cd 2+ , do not restore the enzyme under the conditions studied.Manganese peroxidase (MnP) 1 is an extracellular heme protein produced by virtually all lignin-degrading white-rot fungi (1-3). Sequences have been determined for mnp cDNA (4, 5) and genomic clones (6-8) encoding three MnP isozymes from Phanerochaete chrysosporium, the beststudied lignin-degrading fungus. These sequences and spectroscopic studies of the native and oxidized enzyme intermediates indicate that the heme environment of MnP is similar to that of other plant and fungal peroxidases (4,(9)(10)(11)(12)(13)(14). Kinetic and spectroscopic studies of the purified enzyme indicate a typical peroxidase reaction catalytic cycle:However, MnP is unique in its ability to efficiently oxidize Mn 2+ to Mn 3+ (9,15,16). The released Mn 3+ is stabilized by organic acid chelators, such as oxalate and malonate, which are secreted by the fungus (16, 17). The Mn 3+ -chelator complex is capable of diffusing from the enzyme to oxidize terminal substrates including lignin substructures, phenolic compounds, and pollutants (2, 3 and references therein, 18).The three amino acid residues believed to bind Mn 2+ , D179, E35, and E39, were investigated by site-directed mutagenesis of recombinant MnP homologously expressed in P. chrysosporium (19)(20)(21)(22). These studies, combined with X-ray crystal structure analyses of the proteins (23,24), confirmed that the side-chain carboxylates of D179, E35, and E39 form the only apparent Mn 2+ -binding site in MnP from P. chry...

show abstract

Section: Discussionmentioning

confidence: 67%

Section: Discussionmentioning

confidence: 85%

Formation of a Bis(histidyl) Heme Iron Complex in Manganese Peroxidase at High pH and Restoration of the Native Enzyme Structure by Calcium

et al. 2000

View full text Add to dashboard Cite

show abstract

On Katsuko Kakinuma: Spectroscopic Studies of Redox Centers in NADPH Oxidase – “Identifying and Observing the Key Players that Pass an Electron to Oxygen”

Fujii

Yoshida

2023

NADPH Oxidases Revisited: From Function to Structure

View full text Add to dashboard Cite

Genetics and immunopathology of chronic granulomatous disease

Stasia

2008

Semin Immunopathol

130

View full text Add to dashboard Cite

Chronic granulomatous disease (CGD) is a primary immunodeficiency syndrome characterized by a greatly increased susceptibility to severe fungal and bacterial infections. CGD results from a failure of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme in the patient's phagocytes to produce superoxide. It is caused by mutations in any of four genes that encode the components of the NADPH oxidase. Investigation of CGD patients has identified the different subunits and the genes encoding them. Study of rare CGD variants has highlighted sequences involved in the structural stability of affected components or has provided valuable insights into their function in the oxidase activation mechanism. Functional and molecular CGD diagnosis tests are discussed in this review. Long-term antibiotic prophylaxis has been essential in fighting infections associated with CGD, but approaches based on hematopoietic stem cell transplantation and gene therapy offer great hope for the near future.

show abstract

Spectroscopic identification of the heme axial ligation of cytochrome b₅₅₈ in the NADPH oxidase of porcine neutrophils

Cited by 14 publications

References 32 publications

Formation of a Bis(histidyl) Heme Iron Complex in Manganese Peroxidase at High pH and Restoration of the Native Enzyme Structure by Calcium

Formation of a Bis(histidyl) Heme Iron Complex in Manganese Peroxidase at High pH and Restoration of the Native Enzyme Structure by Calcium

On Katsuko Kakinuma: Spectroscopic Studies of Redox Centers in NADPH Oxidase – “Identifying and Observing the Key Players that Pass an Electron to Oxygen”

Genetics and immunopathology of chronic granulomatous disease

Contact Info

Product

Resources

About

Spectroscopic identification of the heme axial ligation of cytochrome b558 in the NADPH oxidase of porcine neutrophils

Cited by 14 publications

References 32 publications

Formation of a Bis(histidyl) Heme Iron Complex in Manganese Peroxidase at High pH and Restoration of the Native Enzyme Structure by Calcium

Formation of a Bis(histidyl) Heme Iron Complex in Manganese Peroxidase at High pH and Restoration of the Native Enzyme Structure by Calcium

On Katsuko Kakinuma: Spectroscopic Studies of Redox Centers in NADPH Oxidase – “Identifying and Observing the Key Players that Pass an Electron to Oxygen”

Genetics and immunopathology of chronic granulomatous disease

Contact Info

Product

Resources

About

Spectroscopic identification of the heme axial ligation of cytochrome b₅₅₈ in the NADPH oxidase of porcine neutrophils