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SCO2127 与 BldKB 的相互作用及其与变铅青链霉菌形态分化的碳分解代谢物调控的可能联系。

Interaction of SCO2127 with BldKB and its possible connection to carbon catabolite regulation of morphological differentiation in Streptomyces coelicolor.

机构信息

Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, DF, Mexico.

出版信息

Appl Microbiol Biotechnol. 2011 Feb;89(3):799-806. doi: 10.1007/s00253-010-2905-8. Epub 2010 Oct 5.

Abstract

In Streptomyces coelicolor, the sco2127 gene is located upstream of the gene encoding for glucose kinase. This region restores sensitivity to carbon catabolite repression (CCR) of Streptomyces peucetius var. caesius mutants, resistant to 2-deoxyglucose (Dog(R)). In order to search for the possible mechanisms behind this effect, sco2127 was overexpressed and purified for protein-protein interaction studies. SCO2127 was detected during the late growth phase of S. coelicolor grown in a complex media supplemented with 100 mM glucose. Pull-down assays using crude extracts from S. coelicolor grown in the same media, followed by far-western blotting, allowed detection of two proteins bound to SCO2127. The proteins were identified by MALDI-TOF mass spectrometry as SCO5113 and SCO2582. SCO5113 (BldKB) is a lipoprotein ABC-type permease (∼66 kDa) involved in mycelium differentiation by allowing the transport of the morphogenic oligopeptide Bld261. SCO2582, is a putative membrane metalloendopeptidase (∼44 kDa) of unknown function. In agreement with the possible role of SCO2127 in mycelium differentiation, delayed aerial mycelium septation and sporulation was observed when S. coelicolor A3(2) was grown in the presence of elevated glucose concentrations (100 mM), an effect not seen in a Δ-sco2127 mutant derived from it. We speculate that SCO2127 might represent a key factor in CCR of mycelium differentiation by interacting with BldKB.

摘要

在变铅青链霉菌中, sco2127 基因位于编码葡萄糖激酶的基因上游。该区域恢复了对碳分解代谢物阻遏(CCR)的敏感性,使对 2-脱氧葡萄糖(Dog(R))具有抗性的桔青霉素链霉菌变铅青变种突变体敏感。为了寻找这种效应背后的可能机制,过量表达和纯化 sco2127 用于蛋白质-蛋白质相互作用研究。在含有 100mM 葡萄糖的复杂培养基中生长的变铅青链霉菌的生长后期检测到 SCO2127。使用相同培养基中生长的变铅青链霉菌粗提取物进行下拉测定,然后进行远西方印迹杂交,检测到与 SCO2127 结合的两种蛋白质。通过 MALDI-TOF 质谱鉴定,这两种蛋白质分别为 SCO5113 和 SCO2582。SCO5113(BldKB)是一种脂蛋白 ABC 型透性酶(约 66kDa),通过允许形态发生寡肽 Bld261 的运输,参与菌丝体分化。SCO2582 是一种未知功能的假定膜金属内肽酶(约 44kDa)。与 SCO2127 可能在菌丝体分化中的作用一致,当变铅青链霉菌 A3(2)在升高的葡萄糖浓度(100mM)下生长时,观察到气生菌丝体间隔和孢子形成延迟,而从其衍生的Δ-sco2127 突变体中未观察到这种现象。我们推测 SCO2127 可能通过与 BldKB 相互作用成为 CCR 菌丝体分化的关键因素。

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