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

Recombinant SFD isoforms activate vacuolar proton pumps.

Z Zhou1, S B Peng, B P Crider

  • 1Division of Molecular Transport, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75235, USA.

The Journal of Biological Chemistry
|May 21, 1999
PubMed
Summary

The vacuolar proton pump

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

Magnetic character of the low-energy enhancement in <sup>70</sup>Zn.

Nature·2026
Same author

Beta-Decay Half-Lives beyond ^{54}Ca: A Systematic Survey of Decay Properties Approaching the Neutron Dripline.

Physical review letters·2026
Same author

Universal Effective Charges in the sd and fp Shells.

Physical review letters·2025
Same author

Enhanced production of <sup>60</sup>Fe in massive stars.

Nature communications·2024
Same author

Microsecond Isomer at the N=20 Island of Shape Inversion Observed at FRIB.

Physical review letters·2023
Same author

Statistical (n, <math></math> ) cross section model comparison for short-lived nuclei.

The European physical journal. A, Hadrons and nuclei·2023

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • The vacuolar proton pump (V-ATPase) is crucial for cellular processes, involving cytosolic (V1) and membrane (V0) sectors.
  • The V1 sector includes the Sub Fifty-eight-kDa Doublet (SFD) polypeptides, essential for V-ATPase activity but with uncharacterized individual roles.
  • SFD polypeptides are isoforms generated by alternative splicing.

Purpose of the Study:

  • To investigate the distinct functional roles of the 57-kDa (SFDα) and 50-kDa (SFDβ) isoforms of the SFD subunits.
  • To determine if these isoforms are functionally interchangeable in restoring V-ATPase activity.
  • To explore the structural interactions of SFD subunits within the V-ATPase complex.

Main Methods:

  • Expression of recombinant SFDα and SFDβ proteins in Escherichia coli.
  • Reconstitution of SFD-depleted V-ATPase holoenzyme with recombinant SFD isoforms.
  • Assay of ATPase and proton pumping activities.
  • Analysis of V-ATPase from chromaffin granules.

Main Results:

  • Recombinant SFDα and SFDβ restored ATPase and proton pumping activities to SFD-depleted holoenzyme, indicating functional interchangeability.
  • Native V-ATPase from chromaffin granules contains only the SFDα isoform.
  • Both recombinant SFDα and SFDβ equally restored activity to the depleted enzyme.
  • SFDα and SFDβ interact with both V1 and V0 sectors of the V-ATPase.

Conclusions:

  • SFDα and SFDβ isoforms are functionally interchangeable in restoring V-ATPase activity.
  • Despite interchangeability, native V-ATPases may preferentially incorporate specific isoforms.
  • SFD subunits play critical structural and functional roles in coupling ATP hydrolysis to proton transport.

Related Experiment Videos