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

Tubulin dimer dissociation and proteolytic accessibility.

D L Sackett1, D A Zimmerman, J Wolff

  • 1National Institute of Diabetes, Digestive, and Kidney Diseases, Bethesda, Maryland 20894.

Biochemistry
|March 21, 1989
PubMed
Summary

Tubulin dimer cleavage by subtilisin reveals distinct subunit sites. Dilution studies suggest beta-tubulin monomerization, indicating a dissociation constant of 1.5 X 10(-7) M.

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

Colchicine: an ancient drug with novel applications.

The British journal of dermatology·2017
Same author

EPIDEMIOLOGY OF CARDIOVASCULAR DISEASE. AORTIC AND PERIPHERAL ATHEROSCLEROSIS.

A listing of research in the cardiovascular field·2014
Same author

Why Won't Hypertensive Patients Take Their Medicine?

Canadian family physician Medecin de famille canadien·2011
Same author

Hypertension: The Family Physician's Role.

Canadian family physician Medecin de famille canadien·2011
Same author

The impact of continuing education on the quality of patient care.

Canadian family physician Medecin de famille canadien·2011
Same author

An approach to difficult clinical problems.

Canadian family physician Medecin de famille canadien·2011

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Structure

Background:

  • Tubulin dimer comprises alpha and beta subunits, forming microtubules.
  • Each subunit has distinct protease cleavage sites, including C-terminal sites for subtilisin.

Purpose of the Study:

  • To investigate the kinetics and mechanism of subtilisin cleavage on tubulin dimer.
  • To explore the accessibility of cleavage sites and the potential monomer-dimer equilibrium of tubulin.

Main Methods:

  • Enzymatic cleavage assays using subtilisin, trypsin, and chymotrypsin.
  • Kinetic analysis of cleavage rates and determination of pseudo-first-order rate constants.
  • Investigating the effect of tubulin concentration on cleavage kinetics and reversibility.

Main Results:

  • Subtilisin cleaves a C-terminal site on both alpha and beta tubulin subunits.
  • Cleavage kinetics suggest the beta C-terminal site may be more accessible in monomeric tubulin.
  • Dilution experiments revealed a concentration-dependent rapidly digested fraction, attributed to monomeric beta-tubulin.
  • A dissociation constant of approximately 1.5 X 10(-7) M was determined for beta-tubulin monomerization.

Conclusions:

  • Subtilisin cleavage provides insights into tubulin subunit structure and accessibility.
  • Tubulin exhibits a monomer-dimer equilibrium, with beta-tubulin monomerization playing a role in cleavage kinetics.
  • The findings suggest a dynamic model for tubulin assembly and subunit interactions.

Related Experiment Videos