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化学学报
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化学学报 ›› 2010, Vol. 68 ›› Issue (12): 1167-1173. 上一篇    下一篇

研究论文

肽受体跨膜镶嵌的人工细胞膜的构建: 细菌组氨酸蛋白激酶跨膜区多肽在合成肽脂囊泡上的镶嵌

权春善*,1，马金龙1,2，王雪3，李巍4，范圣第1   

  1. (1大连民族学院生命科学学院 大连 116600)
    (2浙江大学化学工程与生物工程学系 杭州 310027)
    (3中国科学院大连化学物理研究所 大连 116023)
    (4大连理工大学环境与生命学院 大连 116064)
  • 投稿日期:2009-12-14 修回日期:2010-03-23 发布日期:2010-04-01
  • 通讯作者: 权春善 E-mail:mikyeken@dlnu.edu.cn
  • 基金资助:

    生物体系外表达跨膜细胞信号转导机能的超分子研究

Construction of an Artificial Signal Transduction System on Bilayer Vesicles: Insertion of Transmembrane Peptide of Bacterial Histidine Protein Kinase on Bilayer Vesicles

Quan Chunshan*,1 Ma Jinlong 1,2 Wang Xue3 Li Wei4 Fan Shengdi1   

  1. (1 College of Life Science, Life Science, Dalian Nationalities University, Dalian 116600)
    (2 Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027)
    (3 Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023)
    (4 School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian 116064)
  • Received:2009-12-14 Revised:2010-03-23 Published:2010-04-01

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被引次数

7 | 1

选择agr I型金黄色葡萄球菌ATCC 6538组氨酸蛋白激酶AgrC蛋白六个跨膜区中与跨膜信号转导过程最密切的一条肽链为受体, 将此多肽采用超声法镶嵌到人工细胞膜上, 采用透射电镜和原子力显微镜对其形态进行了分析, 发现大部分囊泡及镶嵌体系的单个形态为均一球状, 直径大小在50～200 nm范围内. 考察悬浮液pH、体系温度及浓度等因素对其稳定性的影响, 发现在pH为7.0, 温度不超过30 ℃, 多肽和N＋C5Ala2C16的物质的量之比为1∶100时, 镶嵌体系的稳定性最强. 同时, 通过圆二色谱、差示扫描量热分析等手段对镶嵌模型进行表征, 分析发现多肽分子能稳定地插入双层膜囊泡中, 且能主动地插入形成镶嵌体系.

关键词: 跨膜转导, 金黄色葡萄球菌, AgrC, 跨膜镶嵌

One of the six transmembrane domains of AgrC from Staphylococcus aureus ATCC 6538 (agr I type) which is closest with the process of transmembrane signal transduction in quorum sensing was selected as a receptor. This polypeptide was inserted into the artificial cell membrane by ultrasonic procedure, and the pattern of the vesicles was analyzed by transmission electron microscopy (TEM) and atomic force microscopy (AFM). It was found that the single shape of most vesicles (blank and embedded with peptide) was uniform sphere, and the diameter size is from 50 to 200 nm. Considering the impact of suspension pH, concentration and temperature factors to the stability of the embeded system, it is found that the system was the most stable under the following conditions: pH 7.0, temperature lower than 30 ℃, mole ratio of the polypeptide and N＋C5Ala2C16 being 1∶100. Meanwhile, it is also discovered that polypeptide was capable of inserting into bilayer vesicles stably and positively to form embeded system by circular dichroism spectra and differential scanning calorimetry.

Key words: transmembrane transduction, Staphyloccocus aureus, AgrC, transmembrane insertion