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

Conformational changes in yeast phosphoglycerate kinase upon substrate binding

S J Henderson1, E H Serpersu, B S Gerhardt

  • 1Biology Division, Oak Ridge National Laboratory, TN 37831-8077.

Biophysical Chemistry
|December 1, 1994
PubMed
Summary

Small-angle neutron scattering revealed phosphoglycerate kinase (PGK) undergoes a hinge-bending motion during catalysis. Enzyme conformational changes were observed upon substrate binding, indicating a catalytically relevant ternary complex.

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

Fibrinolysis: strategies to enhance the treatment of acute ischemic stroke.

Journal of thrombosis and haemostasis : JTH·2018
Same author

Robust anti-obesity and metabolic effects of a dual GLP-1/glucagon receptor peptide agonist in rodents and non-human primates.

Diabetes, obesity & metabolism·2016
Same author

Risk management in clinical practice. Part 11. Oral surgery.

British dental journal·2011
Same author

Cloning, overexpression, and purification of aminoglycoside antibiotic nucleotidyltransferase (2'')-Ia: conformational studies with bound substrates.

Biochemistry·2001
Same author

Methylation of histone H3 by coactivator-associated arginine methyltransferase 1.

Biochemistry·2001
Same author

Aminoglycoside antibiotics bound to aminoglycoside-detoxifying enzymes and RNA adopt similar conformations.

Cell biochemistry and biophysics·2001

Area of Science:

  • Biochemistry
  • Structural Biology
  • Enzyme Kinetics

Background:

  • Phosphoglycerate kinase (PGK) is a crucial enzyme in glycolysis.
  • Understanding enzyme conformational changes is key to elucidating catalytic mechanisms.

Purpose of the Study:

  • To investigate the conformational dynamics of PGK in response to substrate binding.
  • To characterize the structure of the catalytically relevant ternary complex using small-angle neutron scattering.

Main Methods:

  • Small-angle neutron scattering (SANS) was employed to measure the radius of gyration (Rg) of PGK.
  • Experiments were conducted in D2O with various substrate combinations, including sulfate, CrATP, and PGA.

Main Results:

  • Native PGK exhibited a radius of gyration (Rg) of 24.0 Å.

Related Experiment Videos

  • Binding of substrates (sulfate, CrATP, PGA) induced a decrease in Rg, indicating conformational changes.
  • The most significant Rg decrease (22.6 Å) was observed for PGK in the presence of PGA, CrATP, and sulfate.

    • Conclusions:

    • The observed decrease in Rg supports a hinge-bending motion mechanism for PGK catalysis.
    • The results provide insights into the structural states of PGK within the ternary complex, even without product formation.
    • The data are consistent with a model where substrate binding induces conformational flexibility essential for catalysis.