Structural analysis of the peptide pheromone receptor PlnB, a histidine protein kinase from Lactobacillus plantarum
Structural analysis of the peptide pheromone receptor PlnB, a histidine protein kinase from Lactobacillus plantarum
复制标题
DOI：
10.1128/jb.185.23.6913-6920.2003
复制
发表时间：
2003-12-01
影响因子：
3.2
通讯作者：
Nes, IF
中科院分区：
文献类型：
--
作者：
Johnsborg, O;Diep, DB;Nes, IF
细胞间通讯在革兰氏阳性菌的几个生理过程的调节中起着关键作用。细胞间的通讯通常由分泌的诱导肽信息素(IPs)介导，当在环境中达到阈值浓度时，IPs特异性地激活同源的膜定位的组氨酸蛋白激酶(HPK)。有趣的是，大多数IP激活的HPKs属于一个不同的亚家族(HPK10)。作为研究信息素介导的HPK10亚家族激活机制的一部分，本工作研究了植物乳杆菌PlnB的膜拓扑结构。利用碱性磷酸酶、β-内酰胺酶和β-半乳糖苷酶报告基因与大肠杆菌和乳杆菌的融合实验表明，PlnB通过7个跨膜片段锚定在细胞膜上。通过HPK10成员之间的结构域切换，证明信息素结合和特异性的决定因素包含在跨膜结构域中。结果还表明，HPK10亚家族成员之间的信号转导机制是保守的，其中最后的跨膜片段显然起着关键作用。
Intercellular communication plays a key role in the regulation of several physiological processes in gram-positive bacteria. Cell-cell communication is often mediated by secreted inducer peptide pheromones (IPs), which upon reaching a threshold concentration in the environment specifically activate a cognate membrane-localized histidine protein kinase (HPK). Interestingly, the majority of IP-activated HPKs fall into one distinct subfamily (HPK10). As part of an effort to study the mechanism underlying pheromone-mediated activation of the HPK10 subfamily, the present work investigated the membrane topology of PlnB from Lactobacillus plantarum. Gene fusion experiments with Escherichia coli and Lactobacillus sakei, using alkaline phosphatase, beta-lactamase, and beta-galactosidase reporter fusions, suggested that PlnB is anchored to the cytoplasmic membrane via seven transmembrane segments. By domain switching between HPK10 members, it was demonstrated that the determinants for pheromone binding and specificity are contained within the transmembrane domain. The results also indicate that the mechanism of signal transduction, in which the final transmembrane segment apparently plays a key role, is conserved between members of the HPK10 subfamily.