Chan S J, San Segundo B, McCormick M B, Steiner D F
Proc Natl Acad Sci U S A. 1986 Oct;83(20):7721-5. doi: 10.1073/pnas.83.20.7721.
Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene.
组织蛋白酶B是一种溶酶体硫醇蛋白酶,可能具有额外的非溶酶体功能。为了进一步研究该酶的结构、生物合成模式和细胞内分选,我们利用从人肝癌和肾噬菌体文库中分离的cDNA克隆,确定了人和小鼠前组织蛋白酶B的完整编码序列。核苷酸序列预测,前组织蛋白酶B的一级结构包含339个氨基酸,其组织方式如下:一个17个残基的NH2末端前肽序列,接着是一个62个残基的前肽区域、成熟(单链)组织蛋白酶B中的254个残基以及COOH末端的一个6个残基的延伸。对三种物种(人、小鼠和大鼠)的组织蛋白酶B前体序列进行比较后发现,前肽之间的同源性相对保守,序列同一性最低为68%。特别地,前肽中两个可能具有功能意义的保守序列包括一个潜在的糖基化位点和第59位存在的单个半胱氨酸。对这三个序列的比较分析还表明,组织蛋白酶B前体的加工是一个多步骤过程,在此过程中可能会产生酶活性中间形式。cDNA克隆的可用性将有助于鉴定可能的活性或非活性中间加工形式,以及对组织蛋白酶B基因转录调控的研究。