Characterization of the cAMP phosphodiesterase domain in plant adenylyl cyclase/cAMP phosphodiesterase CAPE from the liverwort Marchantia polymorpha

Characterization of the cAMP phosphodiesterase domain in plant adenylyl cyclase/cAMP phosphodiesterase CAPE from the liverwort Marchantia polymorpha
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地钱植物腺苷酸环化酶/cAMP 磷酸二酯酶 CAPE 中 cAMP 磷酸二酯酶结构域的表征

DOI:
10.1007/s10265-021-01359-4
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发表时间:
2021
影响因子:
2.8
通讯作者:
Kasahara Masahiro
Kasahara Masahiro
中科院分区:
生物学3区
文献类型:
--
作者:
Hayashida Yuta;Yamamoto Chiaki;Takahashi Fumio;Shibata Aika;Kasahara Masahiro

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环腺苷酸(cAMP)作为第二信使参与多种生理反应的调节。最近,我们确定了cAMP合成/水解酶CAPE,其中包含两个催化结构域腺苷酸环化酶(AC)和cAMP磷酸二酯酶(PDE)从地钱。这里我们刻画了M的PDE整环。polymorpha CAPE(MpCAPE-PDE)中表达的纯化蛋白。杆菌MpCAPE-PDE的Km和Vmax分别为30 µM和5.8 nmol min− 1 mg− 1。进一步研究了二价阳离子对PDE活性的影响,发现Ca 2+可增强PDE活性,提示Ca 2+可能通过调节CAPE的PDE活性参与cAMP信号转导。在所测试的偏二磷酸酶抑制剂中,只有双嘧达莫在高浓度下适度抑制偏二磷酸酶活性约40%。相反,3-异丁基-1-甲基黄嘌呤(IBMX)不抑制PDE活性。
Cyclic AMP (cAMP) acts as a second messenger and is involved in the regulation of various physiological responses. Recently, we identified the cAMP-synthesis/hydrolysis enzyme CAPE, which contains the two catalytic domains adenylyl cyclase (AC) and cAMP phosphodiesterase (PDE) from the liverwort Marchantia polymorpha. Here we characterize the PDE domain of M. polymorpha CAPE (MpCAPE-PDE) using the purified protein expressed in E. coli. The K m and V max of MpCAPE-PDE were 30 µM and 5.8 nmol min− 1 mg− 1, respectively. Further, we investigated the effect of divalent cations on PDE activity and found that Ca 2+ enhanced PDE activity, suggesting that Ca 2+ may be involved in cAMP signaling through the regulation of PDE activity of CAPE. Among the PDE inhibitors tested, only dipyridamole moderately inhibited PDE activity by approximately 40% at high concentrations. Conversely, 3-isobutyl-1-methylxanthine (IBMX) did not inhibit PDE activity.
DOI: 10.1073/pnas.0400050101
发表时间: 2004-03-02
影响因子: 11.1
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Esposito, G;Jaiswal, BS;Gossen, JA
通讯作者: Gossen, JA
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DOI: 10.1126/science.1371017
发表时间: 1992
期刊: Science
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DOI: 10.1074/jbc.274.21.15167
发表时间: 1999
期刊: The Journal of Biological Chemistry
影响因子: --
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