title: Proteomic profiling of cellulase-aid-extracted membrane proteins for functional identification of cellulose synthase complexes and their potential associated- components in cotton fibers.
abstract: Cotton fibers are an excellent model for understanding of cellulose biosynthesis in higher plants. In this study, we determined a high cellulose biosynthesis activity in vitro by optimizing biochemical reaction conditions in cotton fibers. By adding a commercial cellulase enzyme into fibers extraction process, we extracted markedly higher levels of GhCESA1 and GhCESA8 proteins and observed an increase in β-1,4-glucan and β-1,3-glucan products in vitro. LC-MS/MS analysis of anti-GhCESA8-immunoprecipitated proteins showed that 19 proteins could be found in three independent experiments including four CESAs (GhCESA1,2,7,8), five well-known non-CESA proteins, one callose synthase (CALS) and nine novel proteins. Notably, upon the cellulase treatment, four CESAs, one CALS and four novel proteins were measured at relatively higher levels by calculating total peptide counts and distinct peptide numbers, indicating that the cellulase-aid-extracted proteins most likely contribute to the increase in β-glucan products in vitro. These results suggest that the cellulase treatment may aid to release active cellulose synthases complexes from growing glucan chains and make them more amenable to extraction. To our knowledge, it is the first time report about the functional identification of the potential proteins that were associated with plant cellulose and callose synthases complexes by using the cellulase-aided protein extraction.
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Ao
authors.initials: A
authors.lastname: Li
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Ruyi
authors.initials: R
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Xianliang
authors.initials: X
authors.lastname: Li
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Mingyong
authors.initials: M
authors.lastname: Liu
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Fan
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Guo
authors.affiliation: School of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Luo
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Chen
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Shengqiu
authors.initials: S
authors.lastname: Feng
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Bingrui
authors.initials: B
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Liangcai
authors.initials: L
authors.lastname: Peng
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.|Biomass and Bioenergy Research Centre, Huazhong Agricultural University, Wuhan 430070, China.|College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Xia
mesh_terms: D002480:Cellulase / Q000378:metabolism*; D002853:Chromatography, Liquid; D032561:Cotton Fiber; D005936:Glucans / Q000032:analysis; D003368:Gossypium / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D047071:beta-Glucans / Q000032:analysis
chemical_list: D005936:Glucans; D008565:Membrane Proteins; D010940:Plant Proteins; D047071:beta-Glucans; C040088:1,4-glucan; C033363:beta-1,3-glucan; D002480:Cellulase