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CaM-stimulated Ca2+-ATPase activity on the symbiosome membrane from broad bean root nodules and its sensitivity to chlorpromazine

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Abstract

Relatively recently, we provided preliminary evidence for stimulating action of CaM on the activity of the symbiosome membrane (SM) Ca2+-ATPase from broad bean root nodules functioning, as to date found by us, as ATP-driven Ca2+/nH+ exchanger. In the present work, CaM effects on the enzyme studied in much more detail by following its catalytic and transport activities. It was found that the kinetics of both ITP hydrolysis by the SM vesicles and symbiosomes and two processes involved in the transport cycle of the Ca2+-ATPase, namely, Ca2+ uptake by symbiosomes and development within them of alkaline pH shift, is markedly accelerated in the presence of CaM and inhibited by its antagonist, chlorpromazine (CP). Based on the data obtained, it was concluded that under selected experimental conditions the observed modulation by CaM of the calcium pump activities is largely caused by the increase in Vmax of the enzyme rather than its apparent Km for Ca2+. These results are discussed in the light of the available literature data showing that the mechanism of CaM action on Ca2+-ATPases in eukaryotic cell membranes may be based not only on dissociation in the presence of Ca2+-CaM complex of the CaM-binding auto-inhibitory domain from their active center but also the use of some other mechanisms not involving any significant structural changes of CaM-binding target proteins.

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Abbreviations

AO:

Acridine orange

CaM:

Calmodulin

CP:

Chlorpromazine

CTC:

Chlorotetracycline

SM:

Symbiosome membrane

ITP:

Inosine triphosphate

FCCP:

Carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone

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Acknowledgements

The results were obtained as part of the implementation of the state task of the Ministry of Education and Science of Russia (FFES-2021-0011).

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Authors and Affiliations

  1. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Botanicheskaya st. 35, Moscow, 127276, Russia

    Valeriya V. Krylova, Rosariya Zartdinova & Stanislav F. Izmailov

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  1. Valeriya V. Krylova
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  2. Rosariya Zartdinova
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  3. Stanislav F. Izmailov
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Correspondence to Valeriya V. Krylova.

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Communicated by A. Sojka.

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Krylova, V.V., Zartdinova, R. & Izmailov, S.F. CaM-stimulated Ca2+-ATPase activity on the symbiosome membrane from broad bean root nodules and its sensitivity to chlorpromazine. Acta Physiol Plant 43, 30 (2021). https://doi.org/10.1007/s11738-020-03196-9

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  • DOI: https://doi.org/10.1007/s11738-020-03196-9

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