How do 14‐3‐3 proteins work? – Gatekeeper phosphorylation and the molecular anvil hypothesis

14‐3‐3 proteins were the first signaling molecules to be identified as discrete phosphoserine/threonine binding modules. This family of proteins, which includes seven isotypes in human cells and up to 15 in plants, plays critical roles in cell signaling events that control progress through the cell cycle, transcriptional alterations in response to environmental cues, and programmed cell death. Despite over 30 years of research, distinct roles for most isotypes remain unknown. Though 14‐3‐3 proteins perform different functions for different ligands, general mechanisms of 14‐3‐3 action include changes in activity of bound ligands, altered association of bound ligands with other cellular components, and changes in intracellular localization of 14‐3‐3‐bound cargo. We present a speculative model where binding of 14‐3‐3 to multiple sites on some ligands results in global ligand conformational changes that mediate their biological effects. For these multi‐site ligands, one binding site is likely to function as a ‘gatekeeper’ whose phosphorylation is necessary for 14‐3‐3 binding but may not always be sufficient for full biological activity. If correct, then 14‐3‐3 may prove to be a bona fide phosphodependent signaling chaperone.