Download MendeleyCrystal structure of the high-affinity Na+,K+-ATPase-ouabain complex with Mg2+ bound in the cation binding site.
Laursen, M., Yatime, L., Nissen, P., Fedosova, N.U.(2013) Proc Natl Acad Sci U S A 110: 10958-10963
- PubMed: 23776223
- DOI: https://doi.org/10.1073/pnas.1222308110
- Primary Citation of Related Structures:
4HYT - PubMed Abstract:
The Na(+),K(+)-ATPase maintains electrochemical gradients for Na(+) and K(+) that are critical for animal cells. Cardiotonic steroids (CTSs), widely used in the clinic and recently assigned a role as endogenous regulators of intracellular processes, are highly specific inhibitors of the Na(+),K(+)-ATPase. Here we describe a crystal structure of the phosphorylated pig kidney Na(+),K(+)-ATPase in complex with the CTS representative ouabain, extending to 3.4 ? resolution. The structure provides key details on CTS binding, revealing an extensive hydrogen bonding network formed by the ¦Â-surface of the steroid core of ouabain and the side chains of ¦ÁM1, ¦ÁM2, and ¦ÁM6. Furthermore, the structure reveals that cation transport site II is occupied by Mg(2+), and crystallographic studies indicate that Rb(+) and Mn(2+), but not Na(+), bind to this site. Comparison with the low-affinity [K2]E2-MgF(x)-ouabain structure [Ogawa et al. (2009) Proc Natl Acad Sci USA 106(33):13742-13747) shows that the CTS binding pocket of [Mg]E2P allows deep ouabain binding with possible long-range interactions between its polarized five-membered lactone ring and the Mg(2+). K(+) binding at the same site unwinds a turn of ¦ÁM4, dragging residues Ile318-Val325 toward the cation site and thereby hindering deep ouabain binding. Thus, the structural data establish a basis for the interpretation of the biochemical evidence pointing at direct K(+)-Mg(2+) competition and explain the well-known antagonistic effect of K(+) on CTS binding.
Organizational Affiliation:
Centre for Membrane Pumps in Cells and Disease (PUMPkin), Danish National Research Foundation, DK-8000 Aarhus C, Denmark.
