A gene (sleC) encoding a spore-cortex-lytic enzyme from Clostridium perfringens S40 spores; cloning, sequence analysis and molecular characterization.
A gene (sleC) encoding a spore-cortex-lytic enzyme from Clostridium perfringens S40 spores; cloning, sequence analysis and molecular characterization.
复制标题
编码来自产气荚膜梭菌 S40 孢子的孢子皮质裂解酶的基因 (sleC)；
作者：
Shigeru Miyata;R. Moriyama;Nobuko Miyahara;Shio Makino
抗血清针对31 kDa的孢子-皮层-裂解酶产生，该酶在产气荚膜梭菌S40孢子萌发期间释放。通过SDS-PAGE分离的休眠孢子和营养细胞组分的Western印迹表明，31 kDa的酶是孢子特异性的，并且休眠孢子中的酶作为36 kDa的蛋白质存在，其没有皮层裂解活性。使用合成的寡核苷酸作为杂交探针，将编码31 kDa酶sleC的基因克隆到大肠杆菌中，并确定整个基因的核苷酸序列。在该阅读框中发现36 kDa蛋白的N-末端氨基酸序列，证实36 kDa蛋白是31 kDa酶的前体形式。推导的氨基酸序列表明，31 kDa的酶产生的前体，包括三个部分：一个N-末端prepro-sequence（114个氨基酸残基），pro-sequence（35个氨基酸残基）和一个成熟的酶（289个氨基酸残基）。有人建议，36 kDa的前酶是非共价连接到皮层层的外部，和处理的前体在发芽过程中释放活性酶。
Antiserum was raised against a 31 kDa spore-cortex-lytic enzyme, which is released during germination of Clostridium perfringens S40 spores. Western blotting of dormant spore and vegetative cell fractions separated by SDS-PAGE indicated that the 31 kDa enzyme is spore-specific and that the enzyme in the dormant spore exists as a 36 kDa protein which has no cortex-lytic activity. A gene encoding the 31 kDa enzyme, sleC, was cloned into Escherichia coli using a synthetic oligonucleotide as a hybridization probe and the nucleotide sequence of the entire gene was determined. The N-terminal amino acid sequence of the 36 kDa protein was found in this reading frame, confirming that the 36 kDa protein is a pro-form of the 31 kDa enzyme. The deduced amino acid sequence indicated that the 31 kDa enzyme is produced as a precursor, comprising three portions; an N-terminal prepro-sequence (114 amino acid residues), a pro-sequence (35 amino acid residues) and a mature enzyme (289 amino acid residues). It is suggested that the 36 kDa pro-enzyme is non-covalently attached to the exterior of the cortex layer, and that the proform is processed to release the active enzyme during germination.
DOI：
10.1016/s0021-9258(18)61070-1
发表时间：
1987-07
期刊：
The Journal of biological chemistry
影响因子：
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作者：
P. Matsudaira
通讯作者：
P. Matsudaira