INDEPENDENT SUBSPACE ANALYSIS
INDEPENDENT SUBSPACE ANALYSIS
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独立的子空间分析
DOI：
10.1111/j.1432-1033.1997.t01-1-00373.x
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
J. Chuang
中科院分区：
文献类型：
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作者：
J. Y. Huang;J. Chuang
分离并测序了一段编码肝筋膜虫肝脂素L蛋白酶完整前体的cDNA。用携带完整基因的质粒转化酿酒糖酵母菌表达和分泌功能活性酶。对温度敏感的酵母突变体进行的实验表明，该酶通过酵母分泌途径进行运输。用截断的基因转化酵母，缺乏肽前编码序列和大部分肽前编码序列，不能表达功能活性酶。酵母表达的酶具有与天然酶相同的物理化学特性，包括最适活性pH值、37°C下的稳定性以及裂解明胶和免疫球蛋白的能力。酶动力学数据表明，天然和酵母表达的组织蛋白酶L1对p2位疏水残基的底物具有相似的特异性。这是第一次报道组织蛋白酶L蛋白酶inS的功能表达。不需要使用酵母分泌信号序列的酿酒酵母。
A cDNA encoding the complete precursor of aFasciola hepaticacathepsin L protease was isolated and sequenced. Functionally active enzyme was expressed and secreted bySaccharomyces cerevisiaetransformed with a plasmid carrying the complete gene. Experiments with temperature‐sensitive yeast mutants showed that the enzyme is trafficked through the yeast secretory pathway. Yeast transformed with a truncated gene, which lacked the pre‐peptide‐encoding and most of the pro‐peptide‐encoding sequences, did not express funtionally active enzyme. The yeast‐expressed enzyme exhibited physico‐chemical properties in common with the native enzyme including, pH optimum for activity, stability at 37°C and ability to cleave gelatin and immunoglobulin. Enzyme kinetic data showed that the native and yeast‐expressed cathepsin L1 have similar specificities for substrates with hydrophobic residues in the P2position. This is the first report of the functional expression of a cathepsin L proteinase inS. cerevisiaethat did not require the use of yeast secretory signal sequences.