Molecular cloning of a calmodulin-dependent phosphatase from murine testis: identification of a developmentally expressed nonneural isoenzyme.
Molecular cloning of a calmodulin-dependent phosphatase from murine testis: identification of a developmentally expressed nonneural isoenzyme.
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DOI：
10.1073/pnas.89.2.529
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发表时间：
1992-01
影响因子：
11.1
通讯作者：
Taro Muramatsu;P. Giri;S. Higuchi;Randall L. Kincaid
中科院分区：
文献类型：
--
作者：
Taro Muramatsu;P. Giri;S. Higuchi;Randall L. Kincaid
从小鼠睾丸文库中克隆了钙调素依赖性蛋白磷酸酶催化亚基（CaM-PrP）的一个独特亚型。1964个碱基对的cDNA序列包含一个开放阅读框，编码一个513个氨基酸的蛋白质（Mr约58，706），其预测的等电点（pI 7.1）比脑亚型的等电点（pI 5.6-5.8）碱性得多。推导的氨基酸序列与其他两个小鼠CaM-PrP基因的同源性为77-81%，并显示出不同的Southern印迹杂交模式，表明它来自一个单独的基因（3型）。大量的2800个核苷酸的mRNA转录在睾丸中观察到，而mRNA的物种在大脑中是检测不到的，因此，它似乎很可能，这CaM-PrP代表一种非神经同工酶。睾丸发育过程中的CaM-PrP mRNA的测量显示，在4-6周期间表达急剧增加，与精子发生的后期阶段相关。这些数据表明，这种磷酸酶同工型可能参与生殖细胞功能，并且与可能调节精子运动性的CaM-PrP的鞭毛相关形式的报道一致[Tash，J.S.，克林克斯，马里兰州，Patel，J.，Means，R. L.，克利角B。& Means，A. R.（1988）J. Cell Biol.106，1625-1633]。
A unique isoform of the catalytic subunit of calmodulin-dependent protein phosphatase (CaM-PrP) was cloned from a murine testis library. The cDNA sequence of 1964 base pairs contained an open reading frame encoding a protein of 513 amino acids (Mr approximately 58,706), the predicted isoelectric point of which (pI 7.1) was much more basic than those of brain isoforms (pI 5.6-5.8). The deduced amino acid sequence was 77-81% identical to two other murine CaM-PrP genes and displayed a distinct Southern blot hybridization pattern, indicating that it was derived from a separate gene (type 3). High amounts of a 2800-nucleotide mRNA transcript were observed in testis, whereas mRNA species were not detectable in brain; thus, it seems likely that this CaM-PrP represents a nonneural isoenzyme. Measurements of CaM-PrP mRNA during testicular development showed a dramatic increase in expression during weeks 4-6, correlating with the later stages of spermatogenesis. These data suggest that this phosphatase isoform may be involved in germ-cell function and are consistent with the report of a flagellum-associated form of CaM-PrP that may regulate sperm motility [Tash, J. S., Krinks, M., Patel, J., Means, R. L., Klee, C. B. & Means, A. R. (1988) J. Cell Biol. 106, 1625-1633].