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Osmolarity, a Key Environmental Signal Controlling Expression of Leptospiral Proteins LigA and LigB and the Extracellular Release of LigA

机译:渗透压,控制钩端螺旋体蛋白LigA和LigB的表达以及LigA的细胞外释放的关键环境信号

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摘要

The high-molecular-weight leptospiral immunoglobulin-like repeat (Lig) proteins are expressed only by virulent low-passage forms of pathogenic Leptospira species. We examined the effects of growth phase and environmental signals on the expression, surface exposure, and extracellular release of LigA and LigB. LigA was lost from stationary-phase cells, while LigB expression was maintained. The loss of cell-associated LigA correlated with selective release of a lower-molecular-weight form of LigA into the culture supernatant, while LigB and the outer membrane lipoprotein LipL41 remained associated with cells. Addition of tissue culture medium to leptospiral culture medium induced LigA and LigB expression and caused a substantial increase in released LigA. The sodium chloride component of tissue culture medium was primarily responsible for the enhanced release of LigA. Addition of sodium chloride, potassium chloride, or sodium sulfate to leptospiral medium to physiological osmolarity caused the induction of both cell-associated LigA and LigB, indicating that osmolarity regulates the expression of Lig proteins. Osmotic induction of Lig expression also resulted in enhanced release of LigA and increased surface exposure of LigB, as determined by surface immunofluorescence. Osmolarity appears to be a key environmental signal that controls the expression of LigA and LigB.
机译:高分子量的钩端螺旋体免疫球蛋白样重复(Lig)蛋白仅通过致病性 Leptospira 物种的低通量低表达形式表达。我们检查了生长期和环境信号对LigA和LigB的表达,表面暴露以及细胞外释放的影响。 LigA从固定相细胞中丢失,而LigB表达得以维持。与细胞相关的LigA的丢失与低分子量形式的LigA选择性释放到培养上清液中有关,而LigB和外膜脂蛋白LipL41仍与细胞相关。将组织培养基添加到钩端螺旋体培养基中会诱导LigA和LigB表达,并导致释放的LigA大量增加。组织培养基中的氯化钠成分主要负责提高LigA的释放。向钩端螺旋体培养基中添加氯化钠,氯化钾或硫酸钠可增加生理渗透压,从而诱导与细胞相关的LigA和LigB,这表明渗透压可调节Lig蛋白的表达。通过表面免疫荧光测定,渗透诱导的Lig表达也导致LigA释放增强和LigB表面暴露增加。渗透压似乎是控制LigA和LigB表达的关键环境信号。

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