Cathepsin K: Isolation and characterization of the murine cDNA and genomic sequence, the homologue of the human pycnodysostosis gene
Cathepsin K: Isolation and characterization of the murine cDNA and genomic sequence, the homologue of the human pycnodysostosis gene
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DOI：
10.1006/bmme.1996.0088
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发表时间：
1996-12-01
期刊：
影响因子：
--
通讯作者：
Desnick, RJ
中科院分区：
文献类型：
--
作者：
Gelb, BD;Moissoglu, K;Desnick, RJ
组织蛋白酶K（EC 3.4.22.38）是一种溶酶体半胱氨酸蛋白酶，与骨吸收密切相关。人类组织蛋白酶It基因在破骨细胞中高度表达，基因突变会导致骨性发育不良，这是一种常染色体隐性骨骼发育不良。为了研究组织蛋白酶K在不同物种间的进化亲缘关系，我们分离了小鼠组织蛋白酶K基因。一个小鼠心脏cDNA克隆pMCatKl含有3'未翻译区、成熟的酶编码序列和大部分前肽。利用cDNA末端的5'快速扩增，从小鼠黑素细胞RNA中扩增出该基因的其余部分。该基因包含一个990-bp的开放阅读框，预测一个329个氨基酸的前原多肽。该蛋白的结构包括一个由15个氨基酸组成的前信号区、一个由99个氨基酸组成的前区和一个由215个氨基酸组成的成熟酶。鉴定出两个潜在的n -糖基化位点，一个在前区，一个在成熟酶中。5'未翻译区为135 bp。3'未翻译区长度为470 bp，包含一个9 bp的聚(a)通道，包含两个聚腺苷化信号。小鼠组织蛋白酶K核苷酸和氨基酸序列在前区和成熟酶上与人、兔和鸡的同源物高度保守。从V129基因组文库中分离到小鼠组织蛋白酶K基因，并对其基因组结构和内含子大小进行了表征，发现其外显子的起始ATG位于外显子2，终止TGA位于外显子8，这一基因组组织与其人类同源物高度保守。小鼠组织蛋白酶K cDNA和基因组序列的可用性将有助于通过基因靶向建立小鼠组织蛋白酶K缺乏症模型。(C) 1996学术出版社，Inc.
Cathepsin K(EC 3.4.22.38) is a lysosomal cysteine protease that is strongly implicated in bone resorption. The human cathepsin It gene is highly expressed in osteoclasts and gene mutations cause pycnodysostosis, an autosomal recessive skeletal dysplasia. To investigate the evolutionary relatedness of cathepsin K across species, the mouse cathepsin K gene was isolated. A mouse heart cDNA clone, pMCatKl, contained the 3' untranslated region, mature enzyme coding sequence, and most of the propeptide. The remainder of the gene was amplified from mouse melanocyte RNA using 5' rapid amplification of cDNA ends. The gene contained a 990-bp open reading frame, predicting a 329-amino-acid prepropolypeptide. The structure of the protein included a 15-amino-acid presignal, a 99-amino-acid proregion, and a 215-amino-acid mature enzyme. Two potential N-glycosylation sites were identified, one in the proregion and one in the mature enzyme. The 5' untranslated region was 135 bp. The 3' untranslated region was 470 bp including a 9-bp poly(A) tract and contained two polyadenylation signals. The mouse cathepsin K nucleotide and amino acid sequences were highly con served with the human, rabbit, and chicken homologues across the proregion and mature enzyme. The mouse cathepsin K gene was isolated from an V129 genomic library, and characterization of its genomic structure and intron sizes revealed exons with the initiation ATG in exon 2 and termination TGA in exon 8, a genomic organization that was highly conserved with its human homologue. The availability of the mouse cathepsin K cDNA and genomic sequences will facilitate generation of a mouse model of cathepsin K deficiency by gene targeting. (C) 1996 Academic Press, Inc.