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

Yeast V1-ATPase: affinity purification and structural features by electron microscopy.

Zhenyu Zhang1, Colleen Charsky, Patricia M Kane

  • 1Department of Biochemistry, University of California-Riverside, Riverside, CA 92521, USA.

The Journal of Biological Chemistry
|September 10, 2003
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

A reversible broad-spectrum antiviral targets the human V-ATPase V<sub>O</sub> domain.

Research square·2026
Same author

Mitochondrial protein import stress causes lysosomal damage and progressive tissue atrophy.

EMBO reports·2026
Same author

Early lipid-mediated responses to hyperosmotic stress at the yeast vacuole.

Molecular biology of the cell·2026
Same author

Sulfonium lipid nanoparticles for intranasal mRNA delivery to lung epithelial and immune cells.

Acta biomaterialia·2025
Same author

Early responses to hyperosmotic stress at the yeast vacuole.

bioRxiv : the preprint server for biology·2025
Same author

Interaction of yeast V-ATPase with TLDc protein Rtc5p.

bioRxiv : the preprint server for biology·2025

Researchers purified yeast V1-ATPase and determined its structure using electron microscopy. The resulting 3D model reveals key structural features, showing similarity to bovine V1-ATPase structures.

Area of Science:

  • Biochemistry
  • Structural Biology
  • Molecular Biology

Background:

  • V-type ATPases (V-ATPases) are essential proton pumps involved in various cellular processes.
  • Understanding the structure of V1-ATPase is crucial for elucidating its function and regulation.

Purpose of the Study:

  • To purify and characterize the V1-ATPase from Saccharomyces cerevisiae.
  • To determine the three-dimensional structure of the yeast V1-ATPase complex.

Main Methods:

  • FLAG affinity tag purification of V1-ATPase.
  • Native gel electrophoresis and mass spectrometry for subunit analysis.
  • Electron microscopy (negative staining and cryo-EM) for structural determination.
  • Angular reconstitution technique for 3D model calculation.

Related Experiment Videos

Main Results:

  • Purified V1-ATPase contained subunits ABCDEFGH, with C at substoichiometric levels.
  • Initial specific Ca-ATPase activity was approximately 6 micromol/min/mg.
  • A 25-A resolution 3D model revealed a pseudo-3-fold symmetric arrangement of six densities around a central cavity.
  • The structure of yeast V1-ATPase closely resembles the V1 domain of intact bovine V1V0-ATPase.

Conclusions:

  • The study successfully purified and structurally characterized yeast V1-ATPase.
  • The high-resolution structural model provides insights into the organization of the V1 domain.
  • Structural similarities suggest conserved architecture across different species' V-ATPases.