Mechanistic Insight into the 14‐3‐3 Protein‐Dependent Activation of Yeast Neutral Trehalase Nth1

Abstract

Trehalases are conserved enzymes catalyzing the hydrolysis of trehalose in a wide range of organisms. The activity of yeast neutral trehalase Nth1 is regulated in PKA, Ca2+ and the 14-3-3 protein-dependent manner. The 14-3-3´s recognize phosphorylated Nth1 and enhance its enzymatic activity through an unknown mechanism [1]. To investigate the structural basis of interaction between Nth1 and 14-3-3, we used HDX-MS, CD spectroscopy, homology modeling and SAXS to identify structural changes occurring upon the complex formation. We showed that the 14-3-3 protein binding changes structure of several regions of phosphorylated Nth1 [2]. The low resolution structural views of Nth1 alone and the Nth1:14-3-3 complex reveal that the 14-3-3 protein binding induces a significant structural rearrangement of the whole Nth1 molecule. The EF-hand-like motif-containing region of Nth1 forms a separate domain that interacts with both the 14-3-3 protein and the catalytic trehalase domain [3]. Our results provide not only a structural view of the 14-3-3-dependent activation of Nth1, but also a bona fide example of the 14-3-3-dependent regulation of other enzyme´s activity. This work was supported by the Czech Science Foundation (Project P207/11/0455).