In Search of Monocot Phosphodiesterases: Identification of a Calmodulin Stimulated Phosphodiesterase from Brachypodium distachyon.
In Search of Monocot Phosphodiesterases: Identification of a Calmodulin Stimulated Phosphodiesterase from Brachypodium distachyon.
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DOI：
10.3390/ijms22179654
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发表时间：
2021-09-06
影响因子：
5.6
通讯作者：
Jaworski K
中科院分区：
文献类型：
--
作者：
Kwiatkowski M;Wong A;Kozakiewicz-Piekarz A;Gehring C;Jaworski K
在植物中，对环境信号的快速和可逆的生物反应可能需要复杂的细胞重新编程。这是通过环核苷酸单磷酸盐(CNMPs)、cAMP和cGMP等信号分子以及钙离子来实现的。虽然cAMP和cGMP在植物中的作用和合成越来越清楚，但cNMP降解酶-磷酸二酯酶(PDE)提供的“关闭信号”却鲜为人知，特别是在单子叶植物中。在本研究中，我们从单子叶短柱海鞘(Brachypodium disachyon，BDPDE1)中筛选出一个候选的PDE，结果表明它能在体外将cNMP水解为5‘NMPs，但对cAMP的选择性高于cGMP。值得注意的是，只有在钙调素(CaM)存在的情况下，才能显著增强PDE的活性，CaM与BDPDE1相互作用，很可能是在预测的CaM结合部位。最后，基于我们的生化、突变和结构分析，我们构建了一个从注释和/或实验验证的跨物种PDE的催化中心提取的全面的氨基酸共识序列，使该搜索基序能够广泛应用于真核生物和原核生物中类似的活性部位的鉴定。
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.
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DOI：
10.1186/1478-811x-8-15
发表时间：
2010-06-25
期刊：
Cell communication and signaling : CCS
影响因子：
--
作者：
Gehring C
通讯作者：
Gehring C
影响因子：
4.6
作者：
Kasahara M;Suetsugu N;Urano Y;Yamamoto C;Ohmori M;Takada Y;Okuda S;Nishiyama T;Sakayama H;Kohchi T;Takahashi F
通讯作者：
Takahashi F
DOI：
10.1098/rstb.1995.0139
发表时间：
1995-10-30
影响因子：
6.3
作者：
BARNES, SA;QUAGGIO, RB;CHUA, NH
通讯作者：
CHUA, NH
影响因子：
6
作者：
Kwiatkowski M;Wong A;Kozakiewicz A;Gehring C;Jaworski K
通讯作者：
Jaworski K
影响因子：
3
作者：
Pettersen, EF;Goddard, TD;Ferrin, TE
通讯作者：
Ferrin, TE