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Topography and functional information of plasma membrane

Science china(Life sciences) 2008年第2期51卷 95-103页

作者： sun delan1, chen jianmin1, song yanmei1, zhu chuanfeng2, pan gebo2 & wan lijun2 1 key laboratory of photosynthesis and environmental molecular physiology, institute of botany, chinese academy of sciences, beijing 100093. china 2 institute of Chemistry, chinese academy of sciences, beijing 100080, china 1. key laboratory of photosynthesis and environmental molecular physiology Institute of Botany Chinese Academy of Sciences Beijing 100093 China 2. institute of Chemistry Chinese Academy of Sciences Beijing 100080 China

By using atomic force microscope (AFM), the topography and function of the plasmalemma surface of the isolated protoplasts from winter wheat mesophyll cells were observed, and compared with dead protoplasts induced by dehydrating stress. The observational results revealed that the plasma membrane of living protoplasts was in a state of polarization. Lipid layers of different cells and membrane areas exhibited distinct active states. The surfaces of plasma membranes were unequal, and were characterized of regionalisation. In addition, lattice structures were visualized in some regions of the membrane surface. These typical structures were assumed to be lipid molecular complexes, which were measured to be 15.8±0.09 nm in diameter and 1.9±0.3 nm in height. Both two-dimensional and three-dimensional imaging showed that the plasmalemma surfaces of winter wheat protoplasts were covered with numerous protruding particles. In order to determine the chemical nature of the protruding particles, living protoplasts were treated by proteolytic enzyme. Under the effect of enzyme, large particles became relatively looser, resulting that their width was increased and their height decreased. The results demonstrated that these particles were likely to be of protein nature. These protein particles at plasmalemma surface were different in size and unequal in distribution. The diameter of large protein particles ranged from 200 to 440 nm, with a central micropore, and the apparent height of them was found to vary from 12 to 40 nm. The diameter of mid-sized protein particles was between 40―60 nm, and a range of 1.8―5 nm was given for the apparent height of them. As for small protein particles, obtained values were 12―40 nm for their diameter and 0.7―2.2 nm for height. Some invaginated pits were also observed at the plasma membrane. They were formed by the endocytosis of protoplast. Distribution density of them at plasmalemma was about 16 pits per 15 μm2. According to their size, we clas

关键词： atomic force microscope winter wheat protoplast plasma membrane surface membrane protein invaginated pits

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