Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Cytochrome c peroxidase from Methylococcus capsulatus Bath

J A Zahn1, D M Arciero, A B Hooper

  • 1Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, 207 Science Building I, Ames, IA 50011-3211, USA.

Archives of Microbiology
|November 5, 1997
PubMed
Summary

Researchers purified bacterial cytochrome c peroxidase from Methylococcus capsulatus Bath. This enzyme contains two distinct hemes with different redox potentials and exhibits temperature-dependent spin-state transitions.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Plant essential oils synergize various pyrethroid insecticides and antagonize malathion in Aedes aegypti.

Medical and veterinary entomology·2019
Same author

Acute Toxicity and Sublethal Effects of Terpenoids and Essential Oils on the Predator Chrysoperla externa (Neuroptera: Chrysopidae).

Neotropical entomology·2017
Same author

Essential oils enhance the toxicity of permethrin against Aedes aegypti and Anopheles gambiae.

Medical and veterinary entomology·2016
Same author

Chlorpyrifos: ecological risk assessment in North American aquatic environments.

Reviews of environmental contamination and toxicology·2006
Same author

Toxicity of pendimethalin to nontarget soil organisms.

Bulletin of environmental contamination and toxicology·2005
Same author

The impact of supplemental dietary methionine sources on volatile compound concentrations in broiler excreta.

Poultry science·2004

Area of Science:

  • Biochemistry
  • Microbiology
  • Enzymology

Background:

  • Obligate methanotrophs like Methylococcus capsulatus Bath play crucial roles in methane oxidation.
  • Cytochrome c peroxidases are essential enzymes involved in cellular redox processes.
  • Understanding enzyme structure and function is key to elucidating metabolic pathways.

Purpose of the Study:

  • To purify and characterize bacterial cytochrome c peroxidase from Methylococcus capsulatus Bath.
  • To investigate the properties of the enzyme's heme centers, including redox potential and spin state.
  • To determine the kinetic parameters for peroxide reduction using a specific electron donor.

Main Methods:

  • Purification of cytochrome c peroxidase from Methylococcus capsulatus Bath.

Related Experiment Videos

  • Characterization of enzyme properties: subunit molecular mass, isoelectric point, and heme content.
  • Spectroscopic analysis to determine heme spin states and redox potentials at different temperatures.
  • Enzyme kinetics assays using ferrocytochrome c555 as the electron donor to measure KM and Vmax.

    • Main Results:

    • The purified enzyme is a homo-dimer with a subunit molecular mass of 35.8 kDa and an isoelectric point of 4.5.
    • The enzyme possesses two hemes: one high-spin, low-potential (Em7 = -254 mV) and one low-spin, high-potential (Em7 = +432 mV).
    • The low-potential heme center undergoes a spin-state transition upon cooling to cryogenic temperatures.
    • Kinetic parameters for peroxide reduction were determined: KM = 510 +/- 100 nM and Vmax = 425 +/- 22 mol ferrocytochrome c555 oxidized min-1 (mole cytochrome c peroxidase)-1.

    Conclusions:

    • Methylococcus capsulatus Bath harbors a unique bacterial cytochrome c peroxidase with distinct heme properties.
    • The enzyme's spin-state transition suggests conformational flexibility and potential regulatory mechanisms.
    • The characterized kinetic parameters provide insights into the enzyme's role in the methanotrophic electron transport chain.