Induction of non-diapause eggs in Bombyx mori by a trehalase inhibitor

The effect of validoxylamine A (VAA), a potent and specific trehalase inhibitor, on the induction of non-diapause in Bombyx mori was examined. The VAA induced non-diapause eggs and prevented the glucogen accumulation in the eggs. Trehalase activity of the pupal ovary was effectively inhibited by the VAA injection.     

葡萄糖3-脱氢酶的研究进展

葡萄糖3-脱氢酶是一种黄素腺嘌呤二核苷酸（FAD）酶,能氧化葡萄糖上的C-3羟基,将葡萄糖转化为相应的3-酮化合物。叙述了葡萄糖3-脱氢酶的来源、性质、生产及其分离纯化。介绍了葡萄糖3-脱氢酶的基因工程进展,以及该酶在工业、医学诊断上的应用前景。     

Structural basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Background

Trehalases are highly conserved enzymes catalyzing the hydrolysis of trehalose in a wide range of organisms. The activity of yeast neutral trehalase Nth1 is regulated in a 14-3-3- and a calcium-dependent manner. The Bmh proteins (the yeast 14-3-3 isoforms) recognize phosphorylated Nth1 and enhance its enzymatic activity through an unknown mechanism.

Methods

To investigate the structural basis of interaction between Nth1 and Bmh1, we used hydrogen/deuterium exchange coupled to mass spectrometry, circular dichroism spectroscopy and homology modeling to identify structural changes occurring upon the complex formation.

Results

Our results show that the Bmh1 protein binding affects structural properties of several regions of phosphorylated Nth1: the N-terminal segment containing phosphorylation sites responsible for Nth1 binding to Bmh, the region containing the calcium binding domain, and segments surrounding the active site of the catalytic trehalase domain. The complex formation between Bmh1 and phosphorylated Nth1, however, is not accompanied by the change in the secondary structure composition but rather the change in the tertiary structure.

Conclusions

The 14-3-3 protein-dependent activation of Nth1 is based on the structural change of both the calcium binding domain and the catalytic trehalase domain. These changes likely increase the accessibility of the active site, thus resulting in Nth1 activation.

General significance

The results presented here provide a structural view of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1, which might be relevant to understand the process of Nth1 activity regulation as well as the role of the 14-3-3 proteins in the regulation of other enzymes.