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

Bacillus subtilis chorismate mutase is partially diffusion-controlled.

P Mattei1, P Kast, D Hilvert

  • 1Laboratorium für Organische Chemie, Swiss Federal Institute of Technology, Zürich, Switzerland.

European Journal of Biochemistry
|April 2, 1999
PubMed
Summary
This summary is machine-generated.

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

Sex-difference of multifactorial intervention on cardiovascular and mortality risk in DKD: post-hoc analysis of a randomised clinical trial.

Cardiovascular diabetology·2024
Same author

Necrotizing sinusitis.

European annals of otorhinolaryngology, head and neck diseases·2020
Same author

Radiologist performance in the interpretation of contrast enemas performed for Hirschsprung's disease in children >1 year of age.

Clinical radiology·2017
Same author

Reclamation of river dredged sediments polluted by PAHs by co-composting with green waste.

The Science of the total environment·2016
Same author

Protein Nanocontainers from Nonviral Origin: Testing the Mechanics of Artificial and Natural Protein Cages by AFM.

The journal of physical chemistry. B·2016
Same author

International Fusion Materials Irradiation Facility injector acceptance tests at CEA/Saclay: 140 mA/100 keV deuteron beam characterization.

The Review of scientific instruments·2014
Same journal

Comparison of expression patterns and cell adhesion properties of the mouse biliary glycoproteins Bgp1 and Bgp2.

European journal of biochemistry·2020
Same journal

AB 3.1.1.1 (or EC 3.1.1.?).

European journal of biochemistry·2020
Same journal

Cdk5.

European journal of biochemistry·2018
Same journal

Structure of the core oligosaccharide of a rough-type lipopolysaccharide of Pseudomonas syringae pv. phaseolicola.

European journal of biochemistry·2004
Same journal

Monitoring ligand-mediated nuclear receptor-coregulator interactions by noncovalent mass spectrometry.

European journal of biochemistry·2004
Same journal

Solution structure of long neurotoxin NTX-1 from the venom of Naja naja oxiana by 2D-NMR spectroscopy.

European journal of biochemistry·2004
See all related articles

Enzyme viscosity affects substrate binding and product release in Bacillus subtilis chorismate mutase (BsCM). Mutants show altered viscosity effects, indicating diffusion plays a role in the enzyme

Area of Science:

  • Biochemistry
  • Enzymology
  • Protein Dynamics

Background:

  • Chorismate mutase catalyzes a key reaction in aromatic amino acid biosynthesis.
  • Understanding enzyme kinetics and reaction mechanisms is crucial for metabolic pathway analysis.
  • Viscosity can probe the role of diffusion in enzyme-catalyzed reactions.

Purpose of the Study:

  • To investigate the impact of microviscosity on the catalytic activity of Bacillus subtilis chorismate mutase (BsCM).
  • To elucidate the rate-limiting steps in the BsCM-catalyzed rearrangement of chorismate to prephenate.
  • To differentiate between diffusion-limited steps and chemical steps in enzyme catalysis.

Main Methods:

  • Enzyme kinetics assays using varying concentrations of viscosogens (e.g., glycerol).

Related Experiment Videos

  • Characterization of wild-type BsCM and specific mutants (C75A, C75S) under different viscosity conditions.
  • Analysis of steady-state kinetic parameters (kcat, kcat/Km) and substrate/product 'on' rates.

    • Main Results:

    • Wild-type BsCM activity decreased significantly with increasing glycerol concentration, indicating viscosity dependence.
    • BsCM mutants C75A and C75S were insensitive to microviscosity changes, ruling out non-specific viscosogen interactions.
    • Kinetic data suggest diffusion contributes 30-40% to the rate-determining steps, involving substrate binding and product release.

    Conclusions:

    • The catalytic mechanism of BsCM is influenced by microviscosity, primarily affecting substrate binding and product release.
    • Diffusion of substrates and products plays a significant role in the overall reaction rate.
    • The accessibility of the active site influences viscosity sensitivity, as seen in the comparison with E. coli chorismate mutase.