中国农业科学2012，Vol.45Issue(11)：2328-2338,11.DOI:10.3864/j.issn.0578-1752.2012.11.023
巴西橡胶树橡胶粒子蛋白质的16-BAC/SDS-PAGE双向电泳及质谱分析
Rubber Particle Protein Analysis of Hevea brasiliensis by Two Dimensional 16-BAC/SDS-PAGE and Mass Spectrometry
摘要
Abstract
[Objective] The objective of the study is to investigate the protein composition and to identify the unknown proteins of the rubber particles in the latex of a rubber tree (Hevea brastiiensis Mull. Arg.). [Method] The rubber particle proteins were separated using two dimensional 16-BAC/SDS-PAGE. The protein spots were cut from the 2-DE gel, subjected to trypsin digestion, and then analyzed using MALDI-TOF-TOF or mass spectrometry de novo sequencing after 4-sulfophenyl isothiocyanate (SPITC) derivatization of tryptic peptides. [Result] A total of 17 rubber particle proteins were identified through a MALDI/ TOF-TOF analysis, among which a new member of the rubber elongation factor (REF) subfamily, hbREF2, was identified using a method of SPITC based de novo amino acid sequencing, and its partial 3' terminal sequence with 87 amino acid residues was obtained. The amino acid sequence of hbREF2 had a high degree of similarity with that of a well-known REF member (gi3 8122474). In. addition, the complete amino acid sequence of one member in the small rubber particle protein (SRPP) subfamily, SRPP (gi|37622210), was obtained through searching the latex transcriptome ESTs database with peptide sequences. [Conclusion] Rubber particle proteins were effectively separated by a 16-BAC/SDS-PAGE method and characterized by mass spectrometry. The data showed the relative abundances of rubber particle proteins and demonstrated that the major protein components of rubber particles are REF and SRPP isoforms. The global identification of the rubber particle proteins is essential to elucidate the biological functions of the rubber particles in the latex of rubber trees.关键词
巴西橡胶树/橡胶粒子/蛋白质/16-BAC/SDS-PAGE双向电泳/质谱分析Key words
Hevea brasiliensis/ rubber particle/ protein/ 16-BAC/SDS-PAGE two dimensional electrophoresis/ mass spectrometry引用本文复制引用
代龙军,项秋兰,黎瑜,聂智毅,康桂娟,段翠芳,曾日中..巴西橡胶树橡胶粒子蛋白质的16-BAC/SDS-PAGE双向电泳及质谱分析[J].中国农业科学,2012,45(11):2328-2338,11.基金项目
国家自然科学基金项目(30860232)、中央级公益性科研院所基本科研业务费项目(1630022012020) （30860232）
