EF-hand motifs of diacylglycerol kinase α interact intra-molecularly with its C1 domains
| Autor: | Tatsuya Yamamoto, Fumio Sakane, Hiromichi Sakai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Intra-molecular interaction
Mutant Diacylglycerol kinase RVH recoverin homology EF-hand Article GST glutathione S-transferase General Biochemistry Genetics and Molecular Biology Homology (biology) EGFP enhanced green fluorescence protein Tandem repeat Recoverin DG diacylglycerol TF trigger factor CR catalytic region C1 domain lcsh:QH301-705.5 ComputingMethodologies_COMPUTERGRAPHICS chemistry.chemical_classification EFHs EF-hand motifs biology EF hand EGTA ethylene glycol tetraacetic acid Cell biology C1Ds C1 domains DGK diacylglycerol kinase Enzyme chemistry Biochemistry lcsh:Biology (General) biology.protein RVHD RVH domain Calcium |
| Zdroj: | FEBS Open Bio, Vol 4, Iss C, Pp 387-392 (2014) FEBS Open Bio |
| ISSN: | 2211-5463 |
| Popis: | Graphical abstract Highlights • We revealed the intra-molecular interaction of DGKα. • The EF-hand motifs of DGKα directly binds to its C1 domains. • The intra-molecular interaction was negatively regulated by Ca2+. • The intra-molecular interaction is important for the activation mechanism of DGKα. Diacylglycerol kinase (DGK) α, which is activated by Ca2+, contains a recoverin homology (RVH) domain, tandem repeats of two Ca2+-binding EF-hand motifs, two cysteine-rich C1 domains and the catalytic domain. We previously found that a DGKα mutant lacking the RVH domain and EF-hands was constitutively active and that the N-terminal region of DGKα, consisting of the RVH domain and EF-hand motifs, interacted intra-molecularly with the C-terminal region containing the C1 and catalytic domains. In this study, we narrowed down the interaction regions of DGKα. At the C-terminal region, the C1 domains are responsible for the intra-molecular interaction. At the N-terminal region, the EF-hand motifs mainly contribute to the interaction. Moreover, using highly purified EF-hand motifs and C1 domains, we demonstrate that they directly bind to each other. The co-precipitation of these two domains was clearly attenuated by the addition of Ca2+. These results indicate that the Ca2+-induced dissociation of the intra-molecular interaction between the EF-hand motifs and the C1 domains of DGKα is the key event that regulates the activity of the enzyme. |
| Databáze: | OpenAIRE |
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