Airo Alessandro, Chan Suzanne L, Martinez Zak, Platt Manu O, Trent Jonathan D
Department of Geological & Environmental Sciences, Stanford University, Stanford, CA, USA.
Cell Biochem Biophys. 2004;40(3):277-88. doi: 10.1385/CBB:40:3:277.
On the basis of acquired thermotolerance and cryotolerance, the optimal heat shock and cold shock temperatures have been determined for Deinococcus radiodurans. A heat shock at 42 degrees C maximized survival at the lethal temperature of 52 degrees C and a cold shock at 20 degrees C maximized survival after repeated freeze-thawing. Enhanced survival from heat shock was found to be strongly dependent on growth stage, with its greatest effect shortly after phase. Increased synthesis of a total of 67 proteins during heat shock and 42 proteins during cold shock were observed by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) and autoradiography. Eight of the most highly induced heat shock proteins shown by 2D PAGE were identified by MALDI-MS as Hsp20, GroEL, DnaK, SodA, Csp, Protease I, and two proteins of unknown function.
基于获得的耐热性和耐冷性,已确定了嗜放射栖热菌的最佳热休克和冷休克温度。42℃的热休克使在52℃致死温度下的存活率最大化,而20℃的冷休克使反复冻融后的存活率最大化。发现热休克后存活率的提高强烈依赖于生长阶段,在对数期后不久效果最佳。通过二维聚丙烯酰胺凝胶电泳(2D PAGE)和放射自显影观察到,热休克期间总共67种蛋白质以及冷休克期间42种蛋白质的合成增加。2D PAGE显示的8种诱导程度最高的热休克蛋白通过基质辅助激光解吸电离质谱(MALDI-MS)鉴定为Hsp20、GroEL、DnaK、SodA、Csp、蛋白酶I以及两种功能未知的蛋白质。