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

Quantitative continuous assay for hyaluronan synthase.

Joanne C Krupa1, David Shaya, Lianli Chi

  • 1Joint Diseases Laboratory, Shriners Hospital for Children, Montreal, Que., Canada H3G 1A6.

Analytical Biochemistry
|December 19, 2006
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

Deep Learning-Driven Saccharide Online Sequencing for Elucidating the Pathological Alterations of Heparan Sulfate in APAP-Induced Acute Liver Injury.

Analytical chemistry·2026
Same author

Probing Glycosaminoglycan-Protein Interactions: Applications of Surface Plasmon Resonance.

Biosensors·2026
Same author

Discovery of Potent, Selective and Efficacious Aminopyrazole Inhibitors of PLK4.

Journal of medicinal chemistry·2025
Same author

Oral efficacy of structurally defined chondroitin sulfate Co-administered with SNAC in Parkinson's disease.

Carbohydrate polymers·2025
Same author

Structural Domain Analysis of Heparin and Heparan Sulfate Combined With Label-Free Quantitative Proteomics to Elucidate Their Functional Diversity in Liver Cancer and APAP-Induced Liver Injury.

Molecular & cellular proteomics : MCP·2025
Same author

D-cysteine impairs tumour growth by inhibiting cysteine desulfurase NFS1.

Nature metabolism·2025

A new assay quantifies hyaluronan synthase activity from Pasteurella multocida (PmHAS). This method measures enzyme kinetics and aids in enzyme purification, offering a rapid tool for biochemical studies.

Area of Science:

  • Biochemistry
  • Enzymology
  • Microbial Pathogenesis

Background:

  • Hyaluronan synthase (HAS) is crucial for bacterial cell wall integrity and virulence in pathogens like Pasteurella multocida (PmHAS).
  • Quantifying PmHAS activity is essential for understanding its role in infection and for developing targeted inhibitors.
  • Existing methods for enzyme activity assays can be time-consuming or lack sensitivity.

Purpose of the Study:

  • To develop a rapid, continuous, and convenient three-enzyme coupled UV absorption assay for quantifying hyaluronan synthase activity.
  • To characterize the kinetic properties and inhibition of a truncated, soluble form of recombinant PmHAS.

Main Methods:

  • A three-enzyme coupled assay utilizing UV absorption was designed.
  • Enzyme activity was measured by monitoring NADH oxidation, linked to UDP production from PmHAS activity.

Related Experiment Videos

  • Products were characterized using gel electrophoresis with a fluorescently labeled primer.

    • Main Results:

    • The developed assay successfully quantified the glucuronic acid and N-acetylglucosamine transferase activities of PmHAS.
    • A truncated soluble form of recombinant PmHAS (residues 1-703) demonstrated time- and concentration-dependent catalytic activity.
    • The assay allows for determination of kinetic parameters, inhibition constants, and mechanistic insights.

    Conclusions:

    • A robust and efficient UV absorption assay was established for PmHAS activity measurement.
    • This assay is suitable for enzyme purification, kinetic studies, and inhibitor screening.
    • The findings provide a valuable tool for further research into PmHAS function and regulation.