MECHANISM OF BACTERIAL SPORE GERMINATION

本提案描述了分析触发机制的努力 巨大芽孢杆菌QM B1551中的细菌孢子萌发。 最近的数据 表明触发剂如L-脯氨酸引起立体特异性 孢子膜的生物物理性质的变化，如通过 荧光去偏振与1，6-二苯基-1，3，5-己三烯（DPH）作为 一个探测器 DPH迁移率的变化将在来自 孢子经过化学修饰以阻止触发， 触发和孢子与改性脂质。 其他技术如ESR， 还将采用DSC和NMR来关联化合物的物理性质。 具有孢子体内特性的膜。 技术也是 被描述为改变孢子膜在体内的物理性质。 触发位点的鉴定将使用脯氨酸亲和类似物 或通过膜蛋白的乙酸酐修饰， 试图纯化脯氨酸位点，目的是确定 当L-脯氨酸与其在孢子中的位点相互作用时，发生物理变化。

This proposal describes efforts to analyze the mechanism for triggering bacterial spore germination in Bacillus megaterium QM B1551. Recent data indicate that a trigger reagent such as L-proline causes a stereospecific change in the biophysical property of the spore membranes as determined by fluorescence depolarization with 1, 6-diphenyl -1, 3, 5-hexatriene (DPH) as a probe. The change in DPH mobility will be examined in membranes from spores chemically modified to block triggering, spores with mutations for triggering and spores with modified lipids. Other techniques such as ESR, DSC and NMR will also be employed to correlate the physical properties of membranes with the in vivo properties of spores. Techniques are also described to alter the physical properties of spore membranes in vivo. Identification of the trigger site will be with a proline affinity analogue or by acetic anhydride modification of the membrane proteins and we will attempt to purify the proline site with the aim of determining what physical changes occur when L-proline interacts with its site in the spore.