Cloning and expression of a rat placental cDNA encoding a novel cathepsin L-related protein.
Cloning and expression of a rat placental cDNA encoding a novel cathepsin L-related protein.
复制标题
编码新型组织蛋白酶 L 相关蛋白的大鼠胎盘 cDNA 的克隆和表达。
DOI：
10.1002/mrd.1080400203
复制
发表时间：
1995
影响因子：
2.5
通讯作者：
Shiverick,KT
中科院分区：
文献类型：
--
作者：
Conliffe,PR;Ogilvie,S;Simmen,RC;Michel,FJ;Saunders,P;Shiverick,KT
组织蛋白酶 L 是由小鼠胎盘和成纤维细胞产生的主要溶酶体半胱氨酸蛋白酶。本研究表征了大鼠胎盘中表达的新型组织蛋白酶 L 相关 mRNA。对大鼠胎盘文库进行免疫学和核苷酸筛选，鉴定出 6 个阳性克隆，其中最大的 pCLRP-9，长度为 924 个碱基对。所有克隆的组合序列均含有711个核苷酸的开放阅读框、终止密码子、聚腺苷酸化位点和3'非翻译区的197个核苷酸，但缺少5'翻译起始密码子。 pCLRP 核苷酸序列与小鼠、大鼠和人组织蛋白酶 L 的核苷酸序列有 60-64% 的同一性。推导的 pCLRP 氨基酸序列编码 237 个氨基酸，与组织蛋白酶 L 的羧基末端序列对齐，并具有半胱氨酸蛋白酶家族特征的活性位点残基。 Northern印迹分析显示pCLRP与胎盘中表达的1.3kb主要mRNA转录物杂交，但肾脏或肝脏中不表达。相比之下，小鼠组织蛋白酶L原的cDNA与在大鼠肾脏和胎盘中表达的1.7kb转录物杂交。在妊娠晚期，大鼠胎盘 pCLRP mRNA 的稳态水平在第 18 天最高，而小鼠组织蛋白酶 L 的稳态水平在妊娠第 20 天最高。小鼠组织蛋白酶 L 抗血清与大鼠胎盘培养基中分子量为 36,000 至 42,000 的四种蛋白质发生交叉反应，其中两种蛋白质在肾脏中不存在。这些数据表明，大鼠胎盘表达多种组织蛋白酶L型蛋白，这些蛋白可能参与胎盘功能和营养供应。 © 1995 Wiley-Liss, Inc.
Cathepsin L is a major lysosomal cysteine protease produced by mouse placenta and fibroblasts. This study characterizes a novel cathepsin L‐related mRNA expressed in rat placenta. Immunological and nucleotide screening of a rat placental library identified six positive clones, the largest, pCLRP‐9, being 924 base pairs in length. The combined sequences of all the clones contain an open reading frame of 711 nucleotides, a termination codon, a polyadenylation site, and 197 nucleotides of 3′ untranslated region, but lack the 5′ translation initiation codon. The pCLRP nucleotide sequence showed 60–64% identity to those of mouse, rat, and human cathepsin L. The deduced amino acid sequence of pCLRP codes for 237 amino acids, which align with the carboxy‐terminal sequence of cathepsin L and has the active site residues characteristic of the cysteine protease family. Northern blot analysis showed hybridization of pCLRP with a major mRNA transcript of 1.3 kilobases expressed in placenta, but not kidney or liver. In contrast, a cDNA for mouse pro‐cathepsin L hybridized with a transcript of 1.7 kilobases expressed in rat kidney, as well as placenta. During late gestation, steady‐state levels of rat placental pCLRP mRNA were highest on day 18, whereas those of mouse procathepsin L were greatest on day 20 of gestation. Antiserum to mouse cathepsin L cross‐reacted with four proteins of molecular weights 36,000 to 42,000 in rat placental culture medium, of which two were absent in the kidney. These data indicate that rat placenta expresses several species of cathepsin L‐type proteins, which may be involved in placental function and nutrient supply. © 1995 Wiley‐Liss, Inc.