LEAF PROTEOME ANALYSIS OF CLEMATIS CHINENSIS: A TRADITIONAL CHINESE MEDICINE (TCM) BY TWO-DIMENSIONAL ELECTROPHORESIS TECHNIQUE

Abstract

Leaf proteome of Clematis chinensis, a traditional Chinese medicine (TCM) was analyzed by two-dimensional electrophoresis (2-DE) technique. The samples were extracted by phenol-SDS method (PSM) with high protein quantity i.e. 2.35±0.345 mg/g (yield/dw). Proteins were visualized by staining of gels by silver stain and CBB. The gel images of each species were compared by Image Master 2D Platinum software for analytical purpose. The 2-DE profile depicted distribution of 1085 spots and out of these only 255 protein spots (23.5%) were common to all analyzed taxa. The visualized protein spots showed pI range from 3.0 to 10.0 (pH) and Mr of 7 kDa to 70 kDa. Twelve proteins were exclusively specific to C. chinensis when compared with its allies, C. finetiana and C. armandii, which may be used as biomarkers. Thirteen proteins were up-regulated in C. finetiana (0.75-0.95 fold) and twelve proteins in C. armandii (1.05-1.66 fold) whilst seven proteins down-regulated (0.66-0.94 fold) in former and three proteins (1.07-1.20 fold) in later one in comparison with C. chinensis. Twenty five differential and similar protein spots were picked and analyzed by LC-MS/MS technique. Identified proteins are related to energy metabolism (ATP synthesis), photosynthesis, nvironmental stimuli, regulating RNA metabolism, growth hormone regulators, evolutionary trends and gene expression. The efficiency and applicability of proteomic approach as biomarker for identification of C. chinensis is discussed in its quality control (QC) perspectives. Leaf proteins of Clematis plants are explored for the first time by 2-DE technique and debated for their metabolic role.