Growth phase-dependent regulation and membrane localization of SpaB, a protein involved in biosynthesis of the lantibiotic subtilin
Growth phase-dependent regulation and membrane localization of SpaB, a protein involved in biosynthesis of the lantibiotic subtilin
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SpaB（一种参与羊毛硫抗生素枯草菌素生物合成的蛋白质）的生长阶段依赖性调节和膜定位
DOI：
10.1128/aem.60.1.1-11.1994
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发表时间：
1994
影响因子：
4.4
通讯作者：
K. Entian
中科院分区：
文献类型：
--
作者：
Z. Gutowski;C. Klein;K. Siegers;K. Bohm;M. Hammelmann;K. Entian
负责羊毛硫抗生素枯草菌素生物合成的信息以spaB基因开始的操纵子样结构组织。spaB基因编码由1，030个氨基酸残基组成的开放阅读框，并且计算出从该基因可以产生理论分子量为120.5kDa的蛋白质。这与十二烷基硫酸钠凝胶电泳和SpaB特异性抗体鉴定后估计的SpaB表观分子量115，000一致。SpaB蛋白与蛋白质EpiB和NisB非常相似，这些蛋白质之前被鉴定为参与表皮素和乳链菌肽的生物合成。从SpaB上游鉴定出特征性的sigma A启动子序列。免疫印迹分析显示SpaB的表达受到强烈调控。没有检测到SpaB蛋白在对数生长期的早期，和最大的SpaB的表达观察到在早期的稳定生长期。SpaB的表达与枯草杆菌素的生物合成密切相关。最近鉴定的两种调控基因spaR和spaK中的任一种的缺失突变，其作为生长期依赖性诱导枯草杆菌素生物合成所必需的“双组分”调控系统（C.克莱因角，澳-地Kaletta和K. D. Entian，Appl. Environ. Microbiol. 59：296-303，1993），也导致SpaB表达失败。为了研究SpaB的细胞内定位，制备了枯草芽孢杆菌囊泡。SpaB蛋白与囊泡组分共沉淀，并且仅在囊泡剧烈重悬后释放。我们的研究结果表明Spa B是膜结合的，枯草杆菌素的生物合成发生在B的细胞质膜上。枯草杆菌。
The information responsible for biosynthesis of the lantibiotic subtilin is organized in an operon-like structure that starts with the spaB gene. The spaB gene encodes an open reading frame consisting of 1,030 amino acid residues, and it was calculated that a protein having a theoretical molecular mass of 120.5 kDa could be produced from this gene. This is consistent with the apparent molecular weight for SpaB of 115,000 which was estimated after sodium dodecyl sulfate-gel electrophoresis and identification with SpaB-specific antibodies. The SpaB protein is very similar to proteins EpiB and NisB, which were identified previously as being involved in epidermin and nisin biosynthesis. Upstream from SpaB a characteristic sigma A promoter sequence was identified. An immunoblot analysis revealed that SpaB expression was strongly regulated. No SpaB protein was detected in the early logarithmic growth phase, and maximum SpaB expression was observed in the early stationary growth phase. The expression of SpaB was strongly correlated with subtilin biosynthesis. Deletion mutations in either of two recently identified regulatory genes, spaR and spaK, which act as a "two-component" regulatory system necessary for growth phase-dependent induction of subtilin biosynthesis (C. Klein, C. Kaletta, and K. D. Entian, Appl. Environ. Microbiol. 59:296-303, 1993), also resulted in failure of SpaB expression. To investigate the intracellular localization of SpaB, vesicles of Bacillus subtilis were prepared. The SpaB protein cosedimented with the vesicle fraction and was released only after vigorous resuspension of the vesicles. Our results suggest that SpaB is membrane associated and that subtilin biosynthesis occurs at the cytoplasmic membrane of B. subtilis.
用于构建trpE融合基因的具有多个克隆位点的高表达载体：pATH载体。
影响因子：
--
作者：
Koerner,TJ;Hill,JE;Myers,AM;Tzagoloff,A
通讯作者：
Tzagoloff,A
两种肠道细菌的生物降解苏氨酸脱水酶的化学表征。
DOI：
10.1016/0167-4838(82)90371-5
发表时间：
1982
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Kim,SS;Datta,P
通讯作者：
Datta,P