title: A protein extraction method for low protein concentration solutions compatible with the proteomic analysis of rubber particles.
abstract: The extraction of high-purity proteins from the washing solution (WS) of rubber particles (also termed latex-producing organelles) from laticifer cells in rubber tree for proteomic analysis is challenging due to the low concentration of proteins in the WS. Recent studies have revealed that proteins in the WS might play crucial roles in natural rubber biosynthesis. To further examine the involvement of these proteins in natural rubber biosynthesis, we designed an efficiency method to extract high-purity WS proteins. We improved our current borax and phenol-based method by adding reextraction steps with phenol (REP) to improve the yield from low protein concentration samples. With this new method, we extracted WS proteins that were suitable for proteomics. Indeed, compared to the original borax and phenol-based method, the REP method improved both the quality and quantity of isolated proteins. By repeatedly extracting from low protein concentration solutions using the same small amount of phenol, the REP method yielded enough protein of sufficiently high-quality from starting samples containing less than 0.02 mg of proteins per milliliter. This method was successfully applied to extract the rubber particle proteins from the WS of natural rubber latex samples. The REP-extracted WS proteins were resolved by 2DE, and 28 proteins were positively identified by MS. This method has the potential to become widely used for the extraction of proteins from low protein concentration solutions for proteomic analysis.
authors.affiliation: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou Hainan, P. R. China.|Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.affiliation: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou Hainan, P. R. China.|Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Tong
authors.affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Lili
authors.initials: L
authors.lastname: Chang
authors.affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Xueru
authors.initials: X
authors.lastname: Meng
authors.affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Limin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou Hainan, P. R. China.
authors.forename: Weimin
authors.initials: W
authors.lastname: Tian
authors.affiliation: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou Hainan, P. R. China.|Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan, P. R. China.
authors.forename: Xuchu
authors.initials: X
authors.lastname: Wang
mesh_terms: D001881:Borates; D005591:Chemical Fractionation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D019800:Phenol; D010940:Plant Proteins / Q000032:analysis* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D012408:Rubber / Q000737:chemistry*
chemical_list: D001881:Borates; D010940:Plant Proteins; C018851:borax; D019800:Phenol; D012408:Rubber
keywords: Mass spectrometry; Plant proteomics; Protein extraction method; Rubber particle; Washing solution