Production of serine protease of Aeromonas sobria is controlled by the protein encoded by the gene lying adjacent to the 3′ end of the protease gene
Production of serine protease of Aeromonas sobria is controlled by the protein encoded by the gene lying adjacent to the 3′ end of the protease gene
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DOI：
10.1111/j.1348-0421.2000.tb02565.x
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发表时间：
2000-01-01
影响因子：
2.6
通讯作者：
Fujii, Y
中科院分区：
文献类型：
--
作者：
Okamoto, K;Nomura, T;Fujii, Y
克隆了温和气单胞菌（Aeromonassobria）蛋白酶基因，测定了其核苷酸序列，该蛋白酶由624个氨基酸残基组成，计算分子量为66，737，7，氨基酸序列显示了细菌丝氨酸蛋白酶的特征。成熟蛋白酶的分子量为65，000道尔顿，氨基酸序列分析表明，成熟蛋白酶前体的氨基末端有一个24个氨基酸的肽段被去除，该肽段可能是跨膜转运的信号肽。随后，我们发现位于蛋白酶基因3'端附近的基因编码的蛋白质（命名为ORF 2蛋白）在蛋白酶的产生中起重要作用，ORF 2基因的突变不影响蛋白酶基因的转录，但导致蛋白酶在细胞中降解，这表明ORF2蛋白是细胞成功生产丝氨酸蛋白酶所必需的。
We cloned a protease gene of Aeromonas sobria and determined its nucleotide sequence, The protease is composed of 624 amino acid residues and its calculated molecular weight is 66,737,7, The amino acid sequence showed the characteristic features of a bacterial serine protease, We expressed the protease gene in Vibrio parahaemolyticus from which the synthesized protease is secreted into the culture medium as the mature form, and purified the mature protease by successive column chromatographies, The size of the mature protease is 65,000 daltons and the amino acid sequence analysis revealed that a 24-amino acid peptide at the amino terminal of the precursor is removed from the mature protease, This peptide might function as a signal peptide in translocation across the inner membrane. Subsequently, we found that the protein, designated ORF2 protein, encoded by the gene lying adjacent to the 3' end of the protease gene plays an important role in production of the protease, Mutation of the ORF2 gene did not affect transcription of the protease gene, but resulted in degradation of the protease in the cell, This shows that ORF2 protein is required for the successful production of the serine protease by cell.