Next Article in Journal
DFT Computed Dielectric Response and THz Spectra of Organic Co-Crystals and Their Constituent Components
Previous Article in Journal
Individual and Combined Effects of Extracellular Polymeric Substances and Whole Cell Components of Chlamydomonas reinhardtii on Silver Nanoparticle Synthesis and Stability
Article Menu
Issue 5 (March-1) cover image

Export Article

Open AccessArticle
Molecules 2019, 24(5), 958; https://doi.org/10.3390/molecules24050958

The Oligomeric State of the Plasma Membrane H+-ATPase from Kluyveromyces lactis

Instituto de Física, Universidad Autónoma de San Luis Potosí; Manuel Nava 6, Zona Universitaria. San Luis Potosí, C.P. 78290. S.L.P., Mexico
*
Author to whom correspondence should be addressed.
Academic Editor: Lajos Novak
Received: 26 January 2019 / Revised: 21 February 2019 / Accepted: 24 February 2019 / Published: 8 March 2019
(This article belongs to the Special Issue Enzyme-Catalyzed Reactions II)
Full-Text   |   PDF [3797 KB, uploaded 13 March 2019]   |  

Abstract

The plasma membrane H+-ATPase was purified from the yeast K. lactis. The oligomeric state of the H+-ATPase is not known. Size exclusion chromatography displayed two macromolecular assembly states (MASs) of different sizes for the solubilized enzyme. Blue native electrophoresis (BN-PAGE) showed the H+-ATPase hexamer in both MASs as the sole/main oligomeric state—in the aggregated and free state. The hexameric state was confirmed in dodecyl maltoside-treated plasma membranes by Western-Blot. Tetramers, dimers, and monomers were present in negligible amounts, thus depicting the oligomerization pathway with the dimer as the oligomerization unit. H+-ATPase kinetics was cooperative (n~1.9), and importantly, in both MASs significant differences were determined in intrinsic fluorescence intensity, nucleotide affinity and Vmax; hence suggesting the large MAS as the activated state of the H+-ATPase. It is concluded that the quaternary structure of the H+-ATPase is the hexamer and that a relationship seems to exist between ATPase function and the aggregation state of the hexamer. View Full-Text
Keywords: H+-ATPase; hexamer; macromolecular assembly; enzyme kinetics; fluorescence; binding site affinity H+-ATPase; hexamer; macromolecular assembly; enzyme kinetics; fluorescence; binding site affinity
Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
SciFeed

Share & Cite This Article

MDPI and ACS Style

Ruiz-Granados, Y.G.; De La Cruz-Torres, V.; Sampedro, J.G. The Oligomeric State of the Plasma Membrane H+-ATPase from Kluyveromyces lactis. Molecules 2019, 24, 958.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top