In Search of Monocot Phosphodiesterases: Identification of a Calmodulin Stimulated Phosphodiesterase from Brachypodium distachyon

International Journal of Molecular Sciences

Aloysius Wong¹  Anna Kozakiewicz-Piekarz²  Christoph Gehring³  Krzysztof Jaworski⁴  Mateusz Kwiatkowski⁴ 

[1]Department of Biology, College of Science and Technology, Wenzhou-Kean University, 88 Daxue Road, Ouhai, Wenzhou 325060, China

[2]Department of Biomedical and Polymer Chemistry, Faculty of Chemistry, Nicolaus Copernicus University, Gagarina St. 7, 87-100 Torun, Poland

[3]Department of Chemistry, Biology and Biotechnology, University of Perugia, Borgo XX giugno, 74, 06121 Perugia, Italy

[4]Department of Plant Physiology and Biotechnology, Nicolaus Copernicus University, Lwowska St. 1, 87-100 Torun, Poland

关键词: phosphodiesterase (PDE);    cAMP;    cGMP;    calmodulin (CaM);    calcium ions;    protein–protein interactions;   

DOI  :  10.3390/ijms22179654

来源: DOAJ

【 摘 要 】

In plants, rapid and reversible biological responses to environmental cues may require complex cellular reprograming. This is enabled by signaling molecules such as the cyclic nucleotide monophosphates (cNMPs) cAMP and cGMP, as well as Ca²⁺. While the roles and synthesis of cAMP and cGMP in plants are increasingly well-characterized, the “off signal” afforded by cNMP-degrading enzymes, the phosphodiesterases (PDEs), is, however, poorly understood, particularly so in monocots. Here, we identified a candidate PDE from the monocot Brachypodium distachyon (BDPDE1) and showed that it can hydrolyze cNMPs to 5′NMPs but with a preference for cAMP over cGMP in vitro. Notably, the PDE activity was significantly enhanced by Ca²⁺ only in the presence of calmodulin (CaM), which interacts with BDPDE1, most likely at a predicted CaM-binding site. Finally, based on our biochemical, mutagenesis and structural analyses, we constructed a comprehensive amino acid consensus sequence extracted from the catalytic centers of annotated and/or experimentally validated PDEs across species to enable a broad application of this search motif for the identification of similar active sites in eukaryotes and prokaryotes.

【 授权许可】

Unknown