Intersubunit bridging by Na+ ions as a rationale for the unusual stability of the c-rings of Na+-translocating F1F0 ATP synthases

Abstract

The oligomeric c-rings of Na+-translocating F1F0 ATP synthases exhibit unusual stability, resisting even boiling in SDS. Here, we show that the molecular basis for this remarkable property is intersubunit crossbridging by Na+ or Li+ ions. The heat stability of c11 was dependent on the presence of Na+ or Li+ ions. For equal stability, 10 times higher Li+ than Na+ concentrations were required, reflecting the 10 times lower binding affinity for Li+ than for Na+. In a recent structural model of c11, the Na+ or Li+ binding ligands are located on neighboring c-subunits, which thus become crossbridged by the binding of either alkali ion with a concomitant increase in the stability of the ring. Site-directed mutagenesis strengthens the essential role of glutamate 65 in the crossbridging of the subunits and also corroborates the proposed stabilizing effect of an ion bridge including aspartate 2.