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

Characterization of folding intermediates using prolyl isomerase

S Veeraraghavan1, B T Nall

  • 1Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760.

Biochemistry
|January 25, 1994
PubMed
Summary

Human peptidyl prolyl cis-trans isomerase (PPI) activity reveals insights into protein folding intermediates. Increased PPI catalytic efficiency suggests a looser protein structure during folding.

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

Occult hepatitis B virus infection among chronic liver disease patients: Insights from a tertiary care hospital.

Indian journal of medical microbiology·2025
Same author

Simultaneous determination of methotrexate, dasatinib and its active metabolite N- deshydroxyethyl dasatinib in rat plasma by LC-MS/MS: method validation and application to pharmacokinetic study.

Arzneimittel-Forschung·2012
Same author

Influence of grapefruit juice on the pharmacokinetics of diltiazem in Wistar rats upon single and multiple dosage regimens.

Die Pharmazie·2009
Same author

Effect of pentoxifylline on the pharmacokinetics of rosiglitazone in Wistar rats.

Methods and findings in experimental and clinical pharmacology·2008
Same author

Genetics of pulmonary fibrosis.

Seminars in respiratory and critical care medicine·2005
Same author

Mapping of the immunodominant T cell epitopes of the protein topoisomerase I.

Annals of the rheumatic diseases·2004

Area of Science:

  • Biochemistry
  • Protein Folding
  • Enzymology

Background:

  • Protein folding is a complex process involving intermediate structures.
  • Peptidyl prolyl cis-trans isomerase (PPI) plays a role in protein folding.
  • Yeast iso-2 cytochrome c undergoes slow folding reactions involving critical proline residues.

Purpose of the Study:

  • To characterize the structure of folding intermediates of yeast iso-2 cytochrome c.
  • To use structure-reactivity relationships of human PPI to probe folding intermediates.
  • To establish the relative catalytic efficiency of PPI as a measure of structure in folding intermediates.

Main Methods:

  • Investigated structure-reactivity relationships of human PPI with yeast iso-2 cytochrome c.
  • Utilized guanidine hydrochloride as a denaturant to perturb protein structure.

Related Experiment Videos

  • Employed site-directed mutagenesis to introduce specific mutations in cytochrome c.
  • Measured the relative catalytic efficiency of PPI ((kcat/Km)rel) as an indicator of structural changes.

    • Main Results:

    • Neither slow folding reaction of wild-type cytochrome c was catalyzed at low denaturant concentrations.
    • Both folding phases were catalyzed by PPI at moderate guanidine hydrochloride concentrations.
    • A mutation in cytochrome c enhanced PPI catalysis of a slow folding phase.
    • Increases in (kcat/Km)rel correlated with protein substrate destabilization.

    Conclusions:

    • The relative catalytic efficiency of PPI serves as a sensitive measure of structure in protein folding intermediates.
    • Destabilization of protein substrates by denaturants or mutation leads to increased PPI accessibility and catalysis.
    • These findings provide insights into the structural dynamics of proline-containing proteins during folding.