Identification of two arginases generated by alternative splicing in the silkworm, Bombyx mori
Identification of two arginases generated by alternative splicing in the silkworm, Bombyx mori
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家蚕中通过选择性剪接产生的两种精氨酸酶的鉴定
DOI：
10.1002/arch.20407
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
K
中科院分区：
文献类型：
--
作者：
Nagaoka;S.;Takata;Y .;Kato;K
精氨酸酶（EC 3.5.3.1）催化精氨酸水解为鸟氨酸和尿素。本研究从家蚕中克隆了两个双磷酸腺苷酶的cDNA。外显子/内含子结构分析表明，命名为bmarg ‐randbmarg‐ f的两个mRNA是由一个基因通过交替使用外显子产生的。据预测，这些bmarg ‐randbmarg‐ f编码几乎相同的氨基酸序列，除了后者有额外的10个N末端残基。重组bmARG-r和bmARG-f在大肠杆菌细胞裂解物中的酶特性大致相似，与使用组织提取物测定的内切酶的酶特性没有显示出大的差异。差异RT-PCR实验和组织分布分析表明，thebmarg-r基因在雄性生殖器官中表达，特别是在乳腺和泡囊中，在交配过程中分泌到精液中并转移到雌性，而thebmarg-f基因在幼虫和成虫的非生殖器官中表达，包括脂肪体和肌肉，其中所产生的酶蛋白被认为留在细胞中。因此，这两种家蚕脱氢酶同工型可能在产物是否从合成产物的细胞中分泌出来方面存在差异。© 2010 Wiley Periodicals，Inc.
Arginase (EC 3.5.3.1) catalyzes the hydrolysis of arginine to ornithine and urea. Here, we have cloned two arginase cDNAs from the silkworm,Bombyx mori. The analysis of exon/intron structures showed that the two mRNAs namedbmarg‐randbmarg‐fwere generated from a single gene by alternative usage of exons. Thebmarg‐randbmarg‐fwere predicted to encode almost the same amino acid sequences, except that the latter had additional ten N‐terminal residues. Recombinant bmARG‐r and bmARG‐f inEscherichia colicell lysates were roughly similar to each other in enzymatic characteristics, which did not show large difference from those of arginases assayed by using tissue extracts. Differential RT‐PCR experiments and tissue distribution analyses of arginase activity indicated that thebmarg‐rgene is expressed in the male reproductive organs, especially in the glandula lacteola and vesicular seminalis, from which it is secreted to the seminal fluid and transferred to the female during copulation, whereas thebmarg‐fgene is expressed in the larval and adult nonreproductive organs including the fat body and muscle, where the produced arginase proteins are considered to stay in the cells. Thus, the two silkworm arginase isoforms may have a difference in whether or not the product is excreted out of the cells in which it is synthesized. © 2010 Wiley Periodicals, Inc.