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

Dynamic requirements for a functional protein hinge.

James G Kempf1, Ju-Yeon Jung, Christina Ragain

  • 1Department of Chemistry, Yale University, P.O. Box 208107, New Haven, CT 06520, USA.

Journal of Molecular Biology
|March 6, 2007
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

Evaluating surface interactions in supported imidazolium ionic liquid phases by using ultrasensitive DNP NMR and DFT studies.

Physical chemistry chemical physics : PCCP·2026
Same author

Facilitating NMR Resonance Assignment with Metabolic Tampering.

bioRxiv : the preprint server for biology·2026
Same author

Kinetic Insights into Cholera Toxin B-Biomimetic Glycan Interactions.

Biomacromolecules·2026
Same author

Exploratory imaging-genetics associations between habenula connectivity and symptom severity in major depressive disorder.

Journal of affective disorders·2026
Same author

Conformational Dynamics and Catalytic Backups in a Hyper-thermostable Engineered Archaeal Protein Tyrosine Phosphatase.

JACS Au·2026
Same author

Comparative Effects of Abdominal Bracing and Valsalva Maneuver on Cerebral and Peripheral Hemodynamics in Healthy Adults: A Randomized Crossover Study.

Medicina (Kaunas, Lithuania)·2025
Same journal

BindRNAgen: Protein-binding RNA sequence generation using latent diffusion models.

Journal of molecular biology·2026
Same journal

Structural basis of HSP90C, a highly active chloroplastic HSP90 chaperone from Arabidopsis thaliana.

Journal of molecular biology·2026
Same journal

Clinical inflammasome biomarkers: Progress and prospects.

Journal of molecular biology·2026
Same journal

Biologically Relevant, Cationic Residues in Human Rhinovirus Stabilize Capsid-Bound RNA Duplexes, and Restrict Capsid Flexibility.

Journal of molecular biology·2026
Same journal

Cryo-EM structures of phage T4 infection intermediate.

Journal of molecular biology·2026
Same journal

A classic fold with a twist: Structural architecture of Dhillonvirus phage Bas18.

Journal of molecular biology·2026
See all related articles

Structural rigidity in enzyme active sites is key for efficient catalysis. This study shows that altering loop flexibility in triosephosphate isomerase (TIM) disrupts substrate binding and catalysis, highlighting the importance of focused motion for protein function.

Area of Science:

  • Enzymology
  • Protein dynamics
  • Structural biology

Background:

  • Triosephosphate isomerase (TIM) is a highly efficient enzyme crucial for glycolysis.
  • Loop 6 in the TIM active site regulates substrate access and catalysis through conformational changes.
  • The N- and C-terminal hinges of loop 6 control its dynamic motion.

Purpose of the Study:

  • To investigate the role of hinge flexibility in loop 6 dynamics and catalytic efficiency of TIM.
  • To compare the solution NMR dynamics of wild-type (WT) TIM with a quintuple mutant (PGG/GGG) with altered hinge regions.
  • To elucidate the relationship between hinge flexibility, loop dynamics, and enzymatic function.

Main Methods:

  • Solution Nuclear Magnetic Resonance (NMR) spectroscopy was employed.

Related Experiment Videos

  • Comparative analysis of WT TIM and a quintuple mutant (PGG/GGG) with glycine substitutions in hinge regions.
  • Analysis of chemical shifts and motional rates to characterize loop 6 dynamics in apo and ligand-bound states.

    • Main Results:

    • The PGG/GGG mutant exhibited significantly slower (<750 s⁻¹) and enhanced nanosecond-timescale loop 6 motions compared to WT TIM.
    • These altered dynamics were observed in both the apo and 2-phosphoglycolate (2-PGA)-bound forms of the mutant.
    • Glycine substitutions reduced the enzyme's response to ligand and induced atypical structural perturbations, leading to conformational heterogeneity.
    • The mutant enzyme showed impaired substrate binding (higher Km) and catalysis (lower kcat).

    Conclusions:

    • Altered hinge flexibility in TIM's loop 6 leads to conformational heterogeneity and reduced catalytic efficiency.
    • Structural rigidity in protein active site hinges is essential for precise control of loop motion and optimal enzyme function.
    • These findings provide insights into the principles of catalytic hinge design in proteins.