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Figure 1 | BMC Complementary and Alternative Medicine

Figure 1

From: Cytotoxic activity of proteins isolated from extracts of Corydalis cava tubers in human cervical carcinoma HeLa cells

Figure 1

Protein purification and electrophoretic analysis. (A) Chromatographic profile of protein purification from C. cava tuber extracts in HT Heparin column (GE Healthcare). Fractions 1 to 9 represented flow-through fractions, 10 to 27 were eluted with a linear NaCl gradient (from 0 to 2 M). The absorbance of all fractions was measured at 280 nm and their DNase activity was estimated using in-gel assay. Proteins present in fractions 16, 17 and 18 were identified using LC-ESI-MS/MS. Fractions 16-19 following three rounds of purification were used in tests on HeLa tumour cell line. (B) In-gel DNase pattern of the gel in which DNase activity of protein fractions originating from C. cava tuber extracts was estimated. ssDNA containing 10% SDS-PAGE, following electrophoresis and incubation (12 h) in 10 mM Ca2+buffer, pH 8.0, was stained with ethidium bromide and viewed under UV light. Nucleolytic activity was noted in fractions Nos.16, 17 and 18 of MW around 30 kDa. Control: purified nucleases from Chelidonium majus milky sap served as positive control. (C) 10% SDS-PAGE following electrophoresis of fraction-contained proteins and silver staining according to Shevchenko et al. [20]. The separated fractions were identical to those in the gel in Fig. 1B. Fractions nos. 16 and 17 each contained 5 protein bands of MW around 30, 32, 35, 38 and 68 kDa, while fraction no. 18 contained an additional band of MW around 140 kDa. The protein bands were numbered 1-6, excised from the gel and sent for identification by mass spectrometry (LC-ESI-MS/MS). M - Protein Molecular Weight Marker (Fermentas).

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