정상 난소 및 난소암조직에서 20 kDa 단백질의 arginine 메칠화 반응에 관한 연구

초록

Al protein methylase I(A1 PMI) activity has been well known to increase in highly proliferative cells, intracellular substrate of which, 20 kDa protein, has been discovered recently. Present study intended to investigate the relevance of ovarian cancer development and arginine methylation of the protein, comparing the intensities of 20 kDa protein and Al protein arginine methylation in normal ovary and ovarian cancer tissues. Normal ovary and ovarian cancer tissues were taken from the same ovarian cancer patient. Al protein expression was induced 30-40 fold by IPTG in BL21 (DES3) LysS E-coli, and Al protein was purified to apparent homogeneity with an yield of 2.89mg per g E-coli. Specific activities of A1 PMI in normal and cancer ovarian tissues were 0.17, 0.2, 1. 18 fold higher in cancer tissue for intracellular substrates, and 5.6, 7.0, 1.25 fold higher in cancer tissue for Al protein. Fluorography revealed methylated 20 kDa protein as an only intracellular substrate of A1 PMI, the intensities of which for normal and cancer ovarian tissues corresponded to enzyme activity measurements. When Al protein was added as exogenous substrates, methylated Al was observed. which parelled the enzyme activities in applied samples, and 20 kDa protein was found to disappear. Conclusively, it was confirmed that 20 kDa protein is the most favored intracellular substrate for AlPMI in physiologic condition. and the protein and Al protein are methylated by the same enzyme competitively.

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