Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (11): 2328-2338.doi: 10.3864/j.issn.0578-1752.2012.11.023

• RESEARCH NOTES • Previous Articles    

Rubber Particle Protein Analysis of Hevea brasiliensis by Two Dimensional 16-BAC/SDS-PAGE and Mass Spectrometry

 DAI  Long-Jun, XIANG  Qiu-Lan, LI  Yu, NIE  Zhi-Yi, KANG  Gui-Juan, DUAN  Cui-Fang, ZENG  Ri-Zhong   

  1. 中国热带农业科学院橡胶研究所/农业部橡胶树生物学与遗传资源利用重点实验室/省部共建国家重点实验室培育基地，海南儋州 571737
  • Received:2011-12-28 Online:2012-06-01 Published:2012-02-21

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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 brasiliensis Müll. 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 (gi|38122474). 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.

Key words: Hevea brasiliensis, rubber particle, protein, 16-BAC/SDS-PAGE two dimensional electrophoresis, mass spectrometry

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