By Colowick S.P., Kaplan N.O.
Liposomes are mobile buildings made of lipid molecules. vital as a mobile version within the examine of easy biology, liposomes also are utilized in medical functions comparable to drug supply and virus reports.
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Vernon, and J. D. Bell, Biochim. Biophys. Acta 1349, 142 (1997). 54 T. Parasassi, G. De Stasio, G. Ravagnan, R. M. Rusch, and E. Gratton, Biophys. J. 60, 179 (1991). 55 C. Balet, K. A. Clingman, and J. Hajdu, Biochem. Biophys. Res. Commun. 150, 561 (1988). 56 F. Radvanyi, L. Jordan, F. Russo-Marie, and C. Bon, Anal. Biochem. 177, 103 (1989). 57 T. Bayburt, B. Z. Yu, I. Street, F. Ghomashchi, F. Laliberte, H. Perrier, Z. Wang, R. Homan, M. K. Jain, and M. H. Gelb, Anal. Biochem. 232, 7 (1995). 58 W.
Weinstein, Biophys. J. 34, 499 (1981). 000 0 100 200 300 400 Seconds Fig. 5. Time course of pyrene-lyso ($2 mol%) excimer-to-monomer fluorescence ratio during hydrolysis of DPPC LUVs (100 M) by sPLA2 (360 nM from A. p. piscivorus venom) at 38 with 10 mM calcium. Excitation was at 344 nm, and emission was at 397 nm (monomer) and 480 nm (excimer). Adapted with permission from Ref. 31. Copyright 1996, Am. Chem. Soc. We have had success with the following procedure. Pyrene-lysophosphatidylcholine is generated by hydrolysis of 1,2-bis-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine (Molecular Probes) with sPLA2.
L. Hazlett and E. A. Dennis, Toxicon 23, 457 (1985). 42  liposomes in the study of PLA2 activity 29 Human. Several human sPLA2 enzymes have been cloned and can be synthesized with bacterial transfection systems followed by folding and purification of the recombinant proteins. 49 Assays of Hydrolysis It is advisable to measure phospholipase activity over time rather than at a single point because the kinetics are rarely linear and are commonly complex. If a single point assay is desirable for screening a large number of compounds as potential inhibitors or for assaying column fractions during purification, it is recommended that either a charged or a micellar substrate be used.