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Updated: Oct 16, 2026

Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis
Published on: June 13, 2021
Epitope-tagged constructs of the yeast plasma-membrane H(+)-ATPase
C F dos Santos1, N D DeWitt, S S Gupta
1Department of Genetics, Yale University School of Medicine, New Haven, CT 06517, USA. tourinho@unisul.rct_sc.br
Abstract:
In this study, two different epitope tags (HA, c-myc) were introduced near the N terminus of the yeast PMA1 H(+)-ATPase. The resulting proteins were indistinguishable from the wild-type ATPase in their ability to travel through the secretory pathway, as judged by quantitative immunoblotting of isolated secretory vesicles. Furthermore, there were no significant abnormalities in ATPase activity (including K(m) for MgATP, Vmax, pH optimum, and IC50 for inhibition by vanadate) or in ATP-dependent proton pumping. Finally, the epitope-tagged ATPases could support normal growth and displayed the expected activation by glucose.
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