Regulatory 14-3-3 proteins bind the atypical motif within the C terminus of the plant plasma membrane H⁺-ATPase via their typical amphipathic groove

Abstract.

The plant plasma membrane H⁺-ATPase contains a C-terminal autoinhibitory domain whose displacement from the catalytic site is caused by binding of regulatory 14-3-3 proteins. Members of the highly conserved 14-3-3 family bind their individual target proteins in a sequence-specific and phosphorylation-dependent manner within a central groove, the latter characterized by the presence of highly invariant residues. However, an atypical binding site for 14-3-3s within the H⁺-ATPase has been identified that does not resemble any other 14-3-3 binding motif. Combination of site-directed mutagenesis with glutathione S-transferase pull-down assays points to the importance of the central 14-3-3 groove for the interaction with the apparently unique site of the H⁺-ATPase. Furthermore, a 14-3-3 dimer is essential for binding such unusual motif.