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

Protein kinase C contains two phorbol ester binding domains.

D J Burns1, R M Bell

  • 1Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.

The Journal of Biological Chemistry
|September 25, 1991
PubMed
Summary

Protein kinase C (PKC) has two distinct phorbol ester binding domains within its regulatory region. These cysteine-rich domains are crucial for phorbol ester binding and may play a role in enzyme regulation.

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

Inter-organ communication: pathways and targets to cardioprotection and neuro-protection. A report from the 12th Hatter Cardiovascular Institute workshop.

Basic research in cardiology·2024
Same author

Remote ischaemic conditioning: defining critical criteria for success-report from the 11th Hatter Cardiovascular Workshop.

Basic research in cardiology·2022
Same author

9th Hatter Biannual Meeting: position document on ischaemia/reperfusion injury, conditioning and the ten commandments of cardioprotection.

Basic research in cardiology·2016
Same author

Cardioprotective Properties of the Platelet P2Y12 Receptor Inhibitor, Cangrelor: Protective in Diabetics and Reliant Upon the Presence of Blood.

Cardiovascular drugs and therapy·2015
Same author

The bizarre imagery effect and intention to learn.

Psychonomic bulletin & review·2013
Same author

Compensator design for improved counterbalancing in high speed atomic force microscopy.

The Review of scientific instruments·2011

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Protein kinase C (PKC) is a family of enzymes involved in various cellular processes.
  • Phorbol esters, such as phorbol dibutyrate (PDBu), are known activators of PKC.
  • Understanding the structural basis of phorbol ester binding to PKC is essential for elucidating its regulatory mechanisms.

Purpose of the Study:

  • To identify the specific domains within protein kinase C (PKC) responsible for phorbol ester binding.
  • To investigate the role of different conserved regions and cysteine-rich sequences in PDBu binding.
  • To characterize the affinity of these domains for phorbol esters.

Main Methods:

  • Expression of various deletion and truncation mutants of PKC using the baculovirus-insect cell system.
  • Binding assays using radiolabeled phorbol dibutyrate ([3H]PDBu) to assess binding activity of mutants.
  • Scatchard analysis to determine the binding affinity of PKC domains for [3H]PDBu.

Main Results:

  • The catalytic domain of PKC did not bind [3H]PDBu, while the regulatory domain did.
  • Deletion of the C1 region, containing cysteine-rich sequences, abolished PDBu binding.
  • Mutants containing either cysteine-rich region exhibited phorbol ester binding activity with affinities comparable to each other but lower than the native enzyme.
  • Peptides from both cysteine-rich regions independently bound [3H]PDBu with high affinity.

Conclusions:

  • Protein kinase C (PKC) possesses at least two distinct phorbol ester binding domains located within its cysteine-rich regions.
  • These cysteine-rich domains are both necessary and sufficient for high-affinity phorbol ester binding.
  • The presence of multiple binding domains suggests a complex regulatory role for phorbol esters in PKC function.

Related Experiment Videos