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从小鼠睾丸中克隆钙调蛋白依赖性磷酸酶:一种在发育过程中表达的非神经同工酶的鉴定。

Molecular cloning of a calmodulin-dependent phosphatase from murine testis: identification of a developmentally expressed nonneural isoenzyme.

作者信息

Muramatsu T, Giri P R, Higuchi S, Kincaid R L

机构信息

Section on Immunology, National Institute on Alcohol Abuse and Alcoholism, Rockville, MD 20852.

出版信息

Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):529-33. doi: 10.1073/pnas.89.2.529.

Abstract

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].

摘要

从鼠睾丸文库中克隆出钙调蛋白依赖性蛋白磷酸酶(CaM-PrP)催化亚基的一种独特同工型。1964个碱基对的cDNA序列包含一个开放阅读框,编码一个由513个氨基酸组成的蛋白质(分子量约为58,706),其预测的等电点(pI 7.1)比脑同工型的等电点(pI 5.6 - 5.8)碱性强得多。推导的氨基酸序列与另外两个鼠CaM-PrP基因的序列有77 - 81%的同一性,并显示出独特的Southern印迹杂交模式,表明它来自一个单独的基因(3型)。在睾丸中观察到大量2800个核苷酸的mRNA转录本,而在脑中未检测到mRNA种类;因此,这种CaM-PrP似乎代表一种非神经同工酶。在睾丸发育过程中对CaM-PrP mRNA的测量显示,在第4 - 6周表达急剧增加,这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,并且与一种可能调节精子运动的与鞭毛相关的CaM-PrP形式的报道一致[Tash, J. S., Krinks, M., Patel, J., Means, R. L., Klee, C. B. & Means, A. R. (1988) J. Cell Biol. 106, 1625 - 1633]。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5675/48272/afab52729c16/pnas01076-0087-a.jpg

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