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

Azotobacter vinelandii citrate synthase

M Rault-Leonardon1, M A Atkinson, C A Slaughter

  • 1Pre-Clinical Science Unit, Department of Veterans Affairs Medical Center, Dallas, Texas 75216.

Biochemistry
|January 10, 1995
PubMed
Summary

We purified Azotobacter vinelandii citrate synthase, revealing a 48,000 Da subunit and hexameric structure, differing from prior research. This enzyme exhibits allosteric properties and NADH inhibition, similar to other Gram-negative bacteria.

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

Histological validation of a type 1 diabetes clinical diagnostic model for classification of diabetes.

Diabetic medicine : a journal of the British Diabetic Association·2020
Same author

Untargeted metabolomic analysis in non-fasted diabetic dogs by UHPLC-HRMS.

Metabolomics : Official journal of the Metabolomic Society·2019
Same author

Raising Awareness: The Need to Promote Allocation of Pancreata From Rare Nondiabetic Donors With Pancreatic Islet Autoimmunity to Type 1 Diabetes Research.

American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons·2016
Same author

Validation of a rapid type 1 diabetes autoantibody screening assay for community-based screening of organ donors to identify subjects at increased risk for the disease.

Clinical and experimental immunology·2016
Same author

The Anatomical Society core regional anatomy syllabus for undergraduate medicine.

Journal of anatomy·2015
Same author

Mobilization without immune depletion fails to restore immunological tolerance or preserve beta cell function in recent onset type 1 diabetes.

Clinical and experimental immunology·2015

Area of Science:

  • Biochemistry
  • Enzymology
  • Microbial Physiology

Background:

  • Citrate synthase is a key enzyme in the Krebs cycle.
  • Previous studies suggested different molecular weights and structures for Azotobacter vinelandii citrate synthase.

Purpose of the Study:

  • To purify and characterize Azotobacter vinelandii citrate synthase.
  • To determine the subunit size, holoenzyme structure, and kinetic properties of the enzyme.
  • To compare these findings with citrate synthases from other organisms.

Main Methods:

  • Enzyme purification techniques.
  • Electrophoresis for subunit size determination.
  • Enzyme activity assays to determine kinetic parameters and allosteric regulation.
  • Amino acid sequencing.

Related Experiment Videos

Main Results:

  • Purified Azotobacter vinelandii citrate synthase has a subunit size of 48,000 Da and a hexameric holoenzyme structure.
  • The enzyme is allosteric with a Hill coefficient of 1.5, showing inhibition by NADH.
  • High ionic strength and AMP alter the Hill coefficient to approximately 1.
  • Partial amino acid sequence shows high similarity to Pseudomonas aeruginosa citrate synthase.

Conclusions:

  • The determined structure and properties of Azotobacter vinelandii citrate synthase differ from previous estimates.
  • The enzyme's allosteric behavior and regulation are comparable to citrate synthases from other Gram-negative, facultative anaerobic organisms.
  • The sequence homology suggests evolutionary relatedness with other bacterial citrate synthases.