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

The putidaredoxin reductase-putidaredoxin electron transfer complex: theoretical and experimental studies.

Vadim Yu Kuznetsov1, Emek Blair, Patrick J Farmer

  • 1Department of Molecular Biology, University of California, Irvine, California 92612-3900, USA.

The Journal of Biological Chemistry
|February 18, 2005
PubMed
Summary

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

An assessment of the cytochrome P450 2-hydroxyisoflavanone synthase (2-HIS) crystal structure.

Communications biology·2026
Same author

Enhancement of Potency and Selectivity of 2-Aminoquinoline-Based Human Neuronal Nitric Oxide Synthase Inhibitors.

Journal of medicinal chemistry·2026
Same author

Potent, Selective, and Brain Penetrant Ether-Linked 2-Aminopyridine Inhibitors of Human Neuronal Nitric Oxide Synthase with Excellent Oral Bioavailability.

Journal of medicinal chemistry·2026
Same author

New Inhibitors of Neuronal Nitric Oxide Synthase for the Treatment of Melanoma.

Journal of medicinal chemistry·2026
Same author

Redox Partner Recognition and Selectivity of Cytochrome P450terp (CYP108A1).

Biochemistry·2025
Same author

Interaction of cytochrome P450 3A4 with the hydrophilic ligand tetraethylene glycol.

Biochemical and biophysical research communications·2025

Electron transfer between putidaredoxin reductase (Pdr) and putidaredoxin (Pdx) was investigated. Bulky residues on Pdx, like Tyr(33), Arg(66), and Trp(106), are crucial for regulating binding affinity and electron transfer rates.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Putidaredoxin reductase (Pdr) and putidaredoxin (Pdx) are key components in bacterial electron transfer pathways.
  • Understanding their interaction is crucial for elucidating enzymatic mechanisms and designing novel biocatalysts.

Purpose of the Study:

  • To investigate the molecular mechanisms of interaction and electron transfer between Pdr and Pdx from Pseudomonas putida.
  • To validate a computer-generated model of the Pdr-Pdx complex through mutagenesis and kinetic studies.

Main Methods:

  • Molecular modeling and computer graphics to generate a Pdr-Pdx complex model.
  • Site-directed mutagenesis of key Pdx residues (Tyr33, Asp38, Arg66, Trp106).
  • Stopped-flow spectroscopy to measure electron transfer rates and binding affinities.

Related Experiment Videos

Main Results:

  • The Pdr-Pdx model showed Pdx docked near Pdr's FAD cofactor, with a 14.6 Å distance to the [2Fe-2S] cluster.
  • Mutations at Tyr(33), Arg(66), and Trp(106) significantly altered electron transfer rates and increased binding affinity to Pdr.
  • Mutant Delta106 Pdx showed similar electron-accepting ability to wild-type, but other mutants exhibited diminished electron transfer from Pdr.

Conclusions:

  • Pdr-Pdx complex formation is driven by steric complementarity, with bulky residues regulating binding.
  • These bulky residues (Tyr33, Arg66, Trp106) facilitate the dissociation of reduced Pdx from Pdr.
  • Electron transfer occurs via multiple pathways, potentially involving Asp(38) and Cys(39), and is not solely dependent on Trp(106).