Korean Journal of Pharmacognosy (생약학회지)

The Korean Society of Pharmacognosy (한국생약학회)
권호 표지

Purification of Phospholipase $A_2$ from Scutellaria baicalensis Suspension Cells

황금 배양 세포로부터 Phospholipase $A_2$의 분리

  • Ma, Choong-Je (Department of Biomaterials Engineering, School of Bioscience and Biotechnology, Kangwon National University) ;
  • Kim, Dae-Kyung (College of Pharmacy, Chungang University)
  • Published : 2009.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

It was previously reported that yeast elicitor transiently increased oleanolic acid and ursolic acid in Scutellaria baicalensis suspension cultures and also doubled phospholipase $A_2$ ($PLA_2$) activity. Thus, $PLA_2$ was purified from the soluble fractions of S. baicalensis suspension cultures and the characters of the purified $PLA_2$ were identified. The $PLA_2$ was purified about 160 times compared with the starting soluble-protein extract from S. baicalensis suspension culture cells. The purified protein showed a molecular mass of about 43 kDa by SDS-PAGE. The purified plant $PLA_2$ had a neutral pH optimum (pH 7.0) and required $Ca^{2+}$ for activity. The $PLA_2$ activity was inhibited by mammalian $PLA_2$ inhibitors such as 5,8,11,14-eicosatetraynoic acid(ETYA) and arachidonyl trifluoromethyl ketone ($AACOCF_3$).

Keywords

References

  1. Divchev, D. and Schieffer, B. (2008) The secretory phospholipase $A_{2}$group IIA: a missing link between inflammation, activated rennin-angiotensin system and atherogenesis? Vascul. Health Risk Manag. 4: 597-604 https://doi.org/10.2147/VHRM.S2008
  2. Dennis, E. A., Rhee, S. G., Billah, M. M. and Hannun, Y. A. (1991) Role of phospholipases in generating lipid second messengers in signal transduction. FASEB J. 5: 2068-2077 https://doi.org/10.1096/fasebj.5.7.1901288
  3. Bergey, D. R., Howe, G. A. and Ryan, C. A. (1996) Polypeptide signaling for plant defensive genes exhibits analogies to defense signaling in animals. Proc. Natl. Acad. Sci. USA 93: 12053-12058 https://doi.org/10.1073/pnas.93.22.12053
  4. Creelman, R. A. and Mullet, J. E. (1997) Biosynthesis and action of jasmonates in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 355-381 https://doi.org/10.1146/annurev.arplant.48.1.355
  5. Ishiguro, S., Kawai-Oda, A., Ueda, J., Nishida, I. and Okada, K. (2001) The defective in anther dehiscence1 gene encodes a novel phospholipase A1 catalyzing the initial step of jasmonic acid biosynthesis, which synchronizes pollen maturation, anther dehiscence, and flower opening in arabidopsis. Plant Cell 13: 2191-2209 https://doi.org/10.1105/tpc.13.10.2191
  6. Ma, C. J. (2008) Cellulase elicitor induced accumulation of capsidiol in Capsicum annumm L. suspension cultures. Biotech. Lett. 30: 961-965 https://doi.org/10.1007/s10529-007-9624-y
  7. Kudo, I., Murakami, M., Hara, S. and Inoue, K. (1993) Mammalian non-pancreatic phospholipases $A_{2}$. Biochim. Biophys. Acta 1170: 217-231 https://doi.org/10.1016/0005-2760(93)90003-R
  8. Munnik, T., Irvine, R. F. and Musgrave, A. (1998) Phospholipid signaling in plants. Biochim. Biophys. Acta 1389: 222-272 https://doi.org/10.1042/BJ20090300
  9. Yoon, H. J., Kim, H. K., Ma, C. J. and Huh, H. (2000) Induced accumulation of triterpenoids in Scutellaria baicalensis suspension cultures using a yeast elicitor. Biotech. Lett. 22: 1071-1075 https://doi.org/10.1023/A:1005610400511
  10. Dole V.P. and Meimertz H. (1960) Microdetermination of long-chain fatty acids in plasma and tissues. J. Biol. Chem. 235: 2595-2599
  11. Jung, K. M. and Kim, D. K. (2000) Purification and characterization of a membrane-associated 48-kilodalton phospholipase $A_{2}$ in leaves of broad bean. Plant Physiol. 123: 1057-1067 https://doi.org/10.1104/pp.123.3.1057
  12. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0
  13. Rordorf, G., Uemura, Y. and Bonventre, J. V. (1991) Characterization of phospholipase $A_{2}$ ($PLA_{2}$) activity in gerbil brain: enhanced activities of cytosolic, mitochondrial, and microsomal forms after ischemia and reperfusion. J. Neurosci. 11: 1829-1836
  14. Scherer, G. F. E. and Arnold, B. (1997) Inhibitors of animal phospholipase A2 enzymes are selective inhibitors of auxin dependent growth: implications for auxin-induced signal transduction. Planta 202: 462-469 https://doi.org/10.1007/s004250050150
  15. Ackermann, E. J., Conde-Frieboes, K. and Dennis, E. A. (1995) Inhibition of macrophage$Ca^{2+}$+-dependent phospholipase $A_{2}$ by bromoenol lactone and trifluoromethyl ketones. J. Biol. Chem. 270: 445-450 https://doi.org/10.1074/jbc.270.1.445
  16. Street, I. P., Lin, H. K., Laliberte, F., Ghomashchi, F., Wang, Z., Perrier, H., Tremblay, N. M., Huang, Z., Weech, P. K. and Gelb, M. H. (1993) Slow- and tight-binding inhibitors of the 85-kDa human phospholipase $A_{2}$. Biochemistry 32: 5935-5940 https://doi.org/10.1021/bi00074a003