Characterization of CoPK02, a Ca2+/calmodulin-dependent protein kinase in mushroom Coprinopsis cinerea.
Characterization of CoPK02, a Ca2+/calmodulin-dependent protein kinase in mushroom Coprinopsis cinerea.
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CoPK02 的表征，CoPK02 是灰霉鬼伞菌中一种 Ca2/钙调蛋白依赖性蛋白激酶。
DOI：
10.1080/09168451.2018.1462692
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Shigeri Y
中科院分区：
文献类型：
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作者：
Yamashita M;Sueyoshi N;Yamada H;Katayama S;Senga Y;Takenaka Y;Ishida A;Kameshita I;Shigeri Y
研究人员从担子菌真菌cinerecoprinopsis的基因组序列中分离出一段与中性曲霉（aspergillus nidulans）中钙调素依赖性蛋白激酶（CaMK） CMKA同源的cDNA。我们将这个编码580个氨基酸，分子量为63,987的序列命名为CoPK02。CoPK02具有12个蛋白激酶特异性亚结构域，分别与大鼠CaMKI、CaMKII、CaMKIV具有43、35、40%的同源性，与CaMK同源基因之一CoPK12具有40%的同源性。在Ca2+/CaM存在下，CoPK02表现出显著的自磷酸化活性和磷酸化外源蛋白。通过CaM-overlay实验，我们证实c端序列（Trp346-Arg358）是钙调素结合位点，并且Ca2+/CaM与CoPK02的结合被CoPK02的自磷酸化所降低。由于CoPK02与CoPK12在不同的进化枝上进化，且与同源于丝裂原活化蛋白激酶活化蛋白激酶的CoPK32基因表达不同，CoPK02和CoPK12可能协同调节Ca2+信号通路。
We surveyed genome sequences from the basidiomycetous mushroomCoprinopsis cinereaand isolated a cDNA homologous to CMKA, a calmodulin-dependent protein kinase (CaMK) inAspergillus nidulans. We designated this sequence, encoding 580 amino acids with a molecular weight of 63,987, as CoPK02. CoPK02 possessed twelve subdomains specific to protein kinases and exhibited 43, 35, 40% identity with rat CaMKI, CaMKII, CaMKIV, respectively, and 40% identity with CoPK12, one of the CaMK orthologs inC. cinerea. CoPK02 showed significant autophosphorylation activity and phosphorylated exogenous proteins in the presence of Ca2+/CaM. By the CaM-overlay assay we confirmed that the C-terminal sequence (Trp346-Arg358) was the calmodulin-binding site, and that the binding of Ca2+/CaM to CoPK02 was reduced by the autophosphorylation of CoPK02. Since CoPK02 evolved in a different clade from CoPK12, and showed different gene expression compared to that of CoPK32, which is homologous to mitogen-activated protein kinase-activated protein kinase, CoPK02 and CoPK12 might cooperatively regulate Ca2+-signaling inC. cinerea.