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

Studies of the M15 beta-galactosidase complementation process

C N Gallagher1, R E Huber

  • 1Division of Biochemistry, Faculty of Science, University of Calgary, Alberta, Canada. huber@acs.ucalgary.ca

Journal of Protein Chemistry
|April 16, 1998
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

Reprint of: Comparison of the Chemical Properties of Selenocysteine and Selenocystine with Their Sulfur Analogs.

Archives of biochemistry and biophysics·2022
Same author

A dentofacial study of male students at the University of Michigan in the physical hardening program.

American journal of orthodontics and oral surgery·2010
Same author

Effect of an E461G mutation of beta-galactosidase (Escherichia coli, lac Z) on pL rate profiles and solvent deuterium isotope effects.

Bioorganic chemistry·2001
Same author

His-391 of beta-galactosidase (Escherichia coli) promotes catalyses by strong interactions with the transition state.

Biochemistry and cell biology = Biochimie et biologie cellulaire·2001
Same author

High resolution refinement of beta-galactosidase in a new crystal form reveals multiple metal-binding sites and provides a structural basis for alpha-complementation.

Protein science : a publication of the Protein Society·2000
Same author

Stabilities of uncomplemented and complemented M15 beta-galactosidase (Escherichia coli) and the relationship to alpha-complementation.

Biochemistry and cell biology = Biochimie et biologie cellulaire·1999

Heat-denatured beta-galactosidase and peptides activate M15 beta-galactosidase. Studies reveal peptide binding to dimer then tetramer formation for full enzyme activity.

Area of Science:

  • Biochemistry
  • Enzymology
  • Protein interactions

Background:

  • Beta-galactosidase is a key enzyme in molecular biology.
  • Understanding enzyme activation and complex formation is crucial for biochemical research.

Purpose of the Study:

  • To investigate the activation mechanisms of M15 beta-galactosidase using various denatured proteins and peptides.
  • To elucidate the stoichiometry and process of peptide binding during enzyme activation.

Main Methods:

  • Native polyacrylamide gel electrophoresis (PAGE) to analyze protein migration and complex formation.
  • Complementation assays using heat-denatured wild-type beta-galactosidase and specific peptides (XP, CB2).
  • Fluorescence-based gel filtration to quantify peptide binding and determine binding stoichiometry.

Related Experiment Videos

Main Results:

  • Heat-denatured wild-type beta-galactosidase, urea, and peptides (XP, CB2) all activated M15 beta-galactosidase.
  • Complementation with XP resulted in three active enzyme forms, while CB2 formed a single active tetrameric form.
  • Peptide binding occurred at a 1:1 ratio (peptide/monomer) for full activity, initially binding to dimer before tetramer formation.

Conclusions:

  • M15 beta-galactosidase can be activated by aggregated or denatured proteins and specific peptides.
  • The N-terminal regions of denatured beta-galactosidase are accessible for complementation.
  • Enzyme activation involves sequential peptide binding, starting with dimer formation and culminating in tetramer assembly at a 1:1 monomer ratio.