STOICHIOMETRY OF THE SODIUM PUMP-PHOSPHOLEMMAN REGULATORY COMPLEX

Seflova, J.; Habibi, N. R.; Yap, J. Q.; Cleary, S. R.; Fang, X.; Kekenes-Huskey, P. M.; Espinoza-Fonseca, L. M.; Bossuyt, J. B.; Robia, S. L.

doi:10.1101/2021.10.12.464104

bioRxiv · 10.1101/2021.10.12.464104

STOICHIOMETRY OF THE SODIUM PUMP-PHOSPHOLEMMAN REGULATORY COMPLEX

Seflova, J.·Habibi, N. R.·Yap, J. Q.·Cleary, S. R.·Fang, X.·Kekenes-Huskey, P. M.·Espinoza-Fonseca, L. M.·Bossuyt, J. B.·Robia, S. L.

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Abstract

The sodium-potassium ATPase (NKA) establishes ion gradients that facilitate many physiological processes. In the heart, NKA activity is regulated by its interaction with phospholemman (PLM, FXYD1). Here we used a novel fluorescence lifetime-based assay to investigate the structure, stoichiometry, and affinity of the NKA-PLM regulatory complex. We observed concentration dependent association of the subunits of NKA-PLM regulatory complex, with avid association of the alpha subunit with the essential beta subunit followed by lower affinity alpha-alpha and alpha-PLM interactions. The data provide the first evidence that the regulatory complex is composed of two alpha subunits associated with two beta subunits, decorated with two PLM regulatory subunits in intact cells. Docking and molecular dynamics simulations generated a structural model of the complex that is consistent with our experimental observations. We propose that alpha-alpha subunit interactions support conformational coupling of the catalytic subunits, which may enhance NKA turnover rate. These observations provide insight into the pathophysiology of heart failure, wherein low NKA expression may be insufficient to support formation of the complete regulatory complex with stoichiometry (alpha-beta-PLM)2.

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Seflova, J., Habibi, N. R., Yap, J. Q., Cleary, S. R., Fang, X., Kekenes-Huskey, P. M., Espinoza-Fonseca, L. M., Bossuyt, J. B., Robia, S. L.. 2021-10-13. STOICHIOMETRY OF THE SODIUM PUMP-PHOSPHOLEMMAN REGULATORY COMPLEX. https://doi.org/10.1101/2021.10.12.464104

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