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当前位置: 首页 > 文章 > 棉花纤维素合酶复合体蛋白的分离与鉴定 棉花学报 2013,25 (2) 129-134
Position: Home > Articles > Characterization of the Cellulose Synthase Complex in Cotton Cotton Science 2013,25 (2) 129-134

棉花纤维素合酶复合体蛋白的分离与鉴定

作  者：
李先良;李傲;彭良才;夏涛
单  位：
华中农业大学作物遗传改良国家重点实验室/生物质与生物能源研究中心/植物科学技术学院/生命科学技术学院
关键词：
棉纤维;纤维素合酶;复合体;次生壁
摘  要：
选用开花后24 d的棉纤维提取原生质膜,用Triton X-100溶解后通过纤维素合酶1(Gossypium hirsutumcellulose synthase1,GhCESA1)抗体进行免疫共沉淀(Co-Immunoprecipitation,Co-IP),以聚丙烯酰胺凝胶电泳(Sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)分离免疫共沉淀的产物,用液相色谱-质谱联用仪(Liquid chromatography-mass spectrometry,LC-MS)进行鉴定。结果表明,棉纤维纤维素合酶复合体可能存在68种蛋白,其中包括8种纤维素合酶(Cellolose synthse,CESA),它们涵盖了棉纤维初生壁和次生壁2种类型纤维素合酶。这说明在次生壁形成的细胞中存在2种类型CESA。本研究也表明复合体中存在非CESA蛋白。
译  名：
Characterization of the Cellulose Synthase Complex in Cotton
作  者：
LI Xian-liang1,2,LI Ao1,PENG Liang-cai1,XIA Tao1 (1.National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research/Biomass and Bioenergy Research Centre/College of Plant Science and Technology/College of Life Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;2.College of Bioengineering,Jingchu University of Technology,Jingmen,Hubei 448000,China)
关键词：
cotton fibers;cellulose synthase;complex;secondary wall
摘  要：
To identify the components of CSCs,plasma membrane(PM) was extracted from cotton fibers at 24 days post anthesis(DPA) and solubilized by triton X-100,from which CSCs were isolated by co-immunoprecipitation mediated by Gossypium hirsutum cellulose synthase1(GhCESA1).The proteins isolated by co-immunoprecipitation mediated by GhCESA1 were characterized by western-blotting using the antibodies of GhCESA1 and Gossypium hirsutum sucrose synthase(GhSuSy).The results show there are GhCESA1 and GhSuSy proteins,which indicates this method can isolate the components of CSCs.Then,CSCs proteins through SDS-PAGE were subjected to nanoflow liquid chromatography-tandem mass spectrometry(LC-MS).68 kinds of proteins were identificated,in which there were 8 kinds of CESA proteins.GhCESA3,5,6 can take part in synthesizing cellulose in promary walls,and the other CESA proteins can be responsible for formation of cellulose in secondary walls.This study suggests that the CSC machinary may be aided by other proteins in addition to cellulose synthase proteins.

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