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Differential Effects of G- and F-Actin on the Plasma Membrane Calcium Pump Activity

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Abstract

We have previously shown that plasma membrane calcium ATPase (PMCA) pump activity is affected by the membrane protein concentration (Vanagas et al., Biochim Biophys Acta 1768:1641–1644, 2007). The results of this study provided evidence for the involvement of the actin cytoskeleton. In this study, we explored the relationship between the polymerization state of actin and its effects on purified PMCA activity. Our results show that PMCA associates with the actin cytoskeleton and this interaction causes a modulation of the catalytic activity involving the phosphorylated intermediate of the pump. The state of actin polymerization determines whether it acts as an activator or an inhibitor of the pump: G-actin and/or short oligomers activate the pump, while F-actin inhibits it. The effects of actin on PMCA are the consequence of direct interaction as demonstrated by immunoblotting and cosedimentation experiments. Taken together, these findings suggest that interactions with actin play a dynamic role in the regulation of PMCA-mediated Ca2+ extrusion through the membrane. Our results provide further evidence of the activation–inhibition phenomenon as a property of many cytoskeleton-associated membrane proteins where the cytoskeleton is no longer restricted to a mechanical function but is dynamically involved in modulating the activity of integral proteins with which it interacts.

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Abbreviations

DMSO:

Dimethyl sulfoxide

EGTA:

Ethylene glycol tetraacetic acid

IOVs:

Inside-out vesicles

MESG:

2-Amino-6-mercapto-7-methylpurine riboside

PMCA:

Plasma membrane calcium ATPase

Tris:

Tris(hydroxymethyl) aminomethane

C12E10 :

Polyoxyethylene glycol monoether with C12 alkyl chain and 10 polyoxyethylene units in the headgroup

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Acknowledgments

The present work was supported by the NIH, Fogarty International Center Grant R03TW006837 to JPFCR and by ANPCYT, CONICET and UBACYT from Argentina. MCDLF, LV, MD, and MFG are doctoral fellows of CONICET. ICM, RCR and JPFCR are established investigators of CONICET, Argentina. EES is an established researcher of Mayo/Clinic Foundation, Rochester, MN, USA.

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Authors and Affiliations

  1. IQUIFIB, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956 (1113), Buenos Aires, Argentina

    Laura Vanagas, María Candelaria de La Fuente, Marianela Dalghi, Mariela Ferreira-Gomes, Rolando C. Rossi, Irene C. Mangialavori & Juan P. F. C. Rossi

  2. Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN, 55905, USA

    Emanuel E. Strehler

Authors
  1. Laura Vanagas
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  2. María Candelaria de La Fuente
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  3. Marianela Dalghi
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  4. Mariela Ferreira-Gomes
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  5. Rolando C. Rossi
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  6. Emanuel E. Strehler
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  7. Irene C. Mangialavori
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  8. Juan P. F. C. Rossi
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Correspondence to Irene C. Mangialavori.

Additional information

Laura Vanagas and María Candelaria de La Fuente are contributed equally to this work.

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Vanagas, L., de La Fuente, M.C., Dalghi, M. et al. Differential Effects of G- and F-Actin on the Plasma Membrane Calcium Pump Activity. Cell Biochem Biophys 66, 187–198 (2013). https://doi.org/10.1007/s12013-012-9467-6

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