Dipartimento di Biologia, Università di Padova, Via Ugo Bassi 58 B, 35121, Padova, Italy.
J Bioenerg Biomembr. 2010 Feb;42(1):37-45. doi: 10.1007/s10863-009-9261-3. Epub 2010 Jan 12.
Thylakoid membranes in higher plant chloroplasts are composed by two distinct domains: stacked grana and stroma lamellae. We developed a procedure for biochemical isolation of grana membranes using mild detergent to maintain membrane structure. Pigment and polypeptide analyses of membrane preparation showed the preparations were indeed enriched in grana membranes. The method was shown to be effective in four different plant species, although with small changes in detergent concentration. Electron microscopy analyses also showed that the preparation consisted of large membrane patches with roughly round shape and diameter comparable with grana membranes in vivo. Furthermore, protein complexes distribution was shown to be maintained with respect to freeze fracture studies, demonstrating that the protocol was successful in isolating membranes close to their in vivo state.
堆叠的基粒和基质片层。我们开发了一种使用温和去污剂分离基粒膜的生化方法，以保持膜结构。膜制剂的色素和多肽分析表明，该制剂确实富含基粒膜。该方法在四种不同的植物物种中均有效，尽管去污剂浓度略有变化。电子显微镜分析还表明，该制剂由大致呈圆形且直径与体内基粒膜相当的大膜片组成。此外，根据冷冻断裂研究，蛋白质复合物的分布也表明得到了维持，这表明该方案成功地分离了接近其体内状态的膜。
