Pruvot G, Cuiné S, Peltier G, Rey P
Département d'Ecophysiologie Végétale et de Microbiologie, Centre d'Etudes de Cadarache, Saint-Paul-lez-Durance, France.
Planta. 1996;198(3):471-9. doi: 10.1007/BF00620065.
Using two-dimensional electrophoresis and Coomassie Blue staining, the accumulation of a 34-kDa protein (named cdsp 34 for chloroplastic drought-induced stress protein) is shown in the thylakoids of Solanum tuberosum plants subjected to a progressive and reversible water deficit. In-vivo labeling experiments showed an increased synthesis of cdsp 34 from the early stages of drought stress (leaf relative water content around 85%) and throughout the constraint. Sequences of the N-terminal part and of four tryptic-digest peptides did not reveal significant homology between the cdsp 34 protein and other known proteins. Western blotting analysis, using a serum raised against the N-terminal part of cdsp 34, confirmed the accumulation of cdsp 34 in thylakoids upon drought stress. From immunoblot analysis of different chloroplastic subfractions, the cdsp 34 protein appears to be an extrinsic protein preferentially located in unstacked stroma thylakoids. Immunoprecipitation of in-vitro-translated products, as well as Southern analysis, showed that the cdsp 34 protein is nuclear encoded. After rewatering of water-stressed plants, the level of cdsp 34 synthesis was reduced, but remained substantially higher than in control plants. Western analysis showed the persistence of a high amount of cdsp 34 in rewatered plants for at least two weeks. Based on the abundance and on the location of cdsp 34 within thylakoids, a putative role for this novel chloroplastic protein is discussed in relation to the tolerance of the photosynthetic apparatus of higher plants to dehydration.
利用二维电泳和考马斯亮蓝染色法,在经历渐进性且可逆水分亏缺的马铃薯植株类囊体中,显示出一种34 kDa蛋白(命名为cdsp 34,即叶绿体干旱诱导应激蛋白)的积累。体内标记实验表明,从干旱胁迫早期阶段(叶片相对含水量约85%)直至整个胁迫过程,cdsp 34的合成均增加。cdsp 34蛋白N端部分及四个胰蛋白酶消化肽段的序列,未显示该蛋白与其他已知蛋白有显著同源性。利用针对cdsp 34 N端部分产生的血清进行的蛋白质印迹分析,证实干旱胁迫下类囊体中cdsp 34的积累。通过对不同叶绿体亚组分的免疫印迹分析,cdsp 34蛋白似乎是一种外在蛋白,优先定位于未堆叠的基质类囊体中。体外翻译产物的免疫沉淀以及Southern分析表明,cdsp 34蛋白是核编码的。水分胁迫植株复水后,cdsp 34的合成水平降低,但仍显著高于对照植株。蛋白质印迹分析表明,复水植株中大量的cdsp 34至少持续存在两周。基于cdsp 34在类囊体中的丰度和定位,讨论了这种新型叶绿体蛋白在高等植物光合机构对脱水耐受性方面的假定作用。