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Ca(2+)-ATPase-driven calcium accumulation in Ustilago maydis plasma membrane vesicles
A Hernández1, D T Cooke, D T Clarkson
1Department of Agricultural Sciences, University of Bristol, AFRC Institute of Arable Crops Research, UK.
Microbiology (Reading, England)
|November 1, 1994
Summary
Calcium transport in Ustilago maydis plasma membrane vesicles is primarily regulated by a P-type ATPase. This energy-dependent process utilizes ATP and Mg2+, showing sensitivity to vanadate and erythrosin B inhibitors.
Area of Science:
- * Biochemistry and Molecular Biology
- * Plant Pathology and Mycology
Background:
- * Understanding calcium (Ca2+) transport mechanisms is crucial for cellular function in fungi.
- * Ustilago maydis, a significant plant pathogen, requires investigation into its ion transport systems.
Purpose of the Study:
- * To characterize the Ca2+ transport system in plasma membrane vesicles of Ustilago maydis.
- * To identify the molecular nature and regulatory properties of this Ca2+ transporter.
Main Methods:
- * Isolation of plasma membrane vesicles from Ustilago maydis cells.
- * Measurement of Ca2+ uptake and release using radiolabeled Ca2+.
- * Assessment of inhibitor effects (vanadate, erythrosin B, ionophores, channel blockers) and energy dependence (ATP, GTP, Mg2+).
Main Results:
- * Ca2+ transport is dependent on Adenosine Triphosphate (ATP) and Magnesium ions (Mg2+).
- * Vanadate and erythrosin B significantly inhibited Ca2+ transport, with an IC50 of approximately 10 microM.
- * Protonophores caused partial inhibition, while specific Ca2+-channel blockers had no effect, suggesting a non-channel mediated transport.
Conclusions:
- * The Ca2+ transport in Ustilago maydis is mediated by a P-type ATPase.
- * This ATPase exhibits properties similar to those identified in higher plants, indicating conserved mechanisms.
- * The findings provide insights into fungal ion homeostasis and potential targets for disease management.