인지질분해효소D에 의해 유도된 소낭 융합에 대한 상 비대칭의 영향 = Phase Asymmetry Effect on Vesicle Fusion Induced by Phospholipase D

다국어 초록 (Multilingual Abstract)

Spherical phospholipid bilayers, vesicles, were formed with respect to phase of each layer via a double emulsion technique. The conversion of phosphatidylcholine (PC) to phosphatidic acid (PA) at the outer layer, caused by phospholipase D (PLD), induced a curvature change in the vesicles, which eventually led them to fuse each other. The effect of the lipid layer physical-properties on the PLD-induced vesicle fusion was investigated using the fluorescence intensity change. 8-Aminonaphthalene-1,3,6-trisulfonic acid disodium salt(ANTS) and p-Xylene-bis(N-pyridinium bromide)(DPX) were encapsulated in the vesicles, respectively, for the quantification of the fusion. The fluorescence scale was calibrated with the fluorescence of a 1/1 mixture of ANTS and DPX vesicles in NaCl buffer taken as 100% fluorescence (0% fusion) and the vesicles containing both ANTS and DPX as 0% fluorescence (100% fusion), considering the leakage into the medium studied directly in a separate experiment using vesicles containing both ANTS and DPX. It was observed that the fusion occurred to the liquid-phase of the inner layer only. The fusion behaviors were very similar for both solid and liquid of the outer layer. However, the leakage was faster for the solid-phase outer-layer than the liquid-phase outerlayer. The difference in the leakage seems to be caused by the lipid concentration and the lateral diffusivity in the layer.

참고문헌 (Reference)

1 함수미, "긴 사슬 지방산 생산을 위해 재조합된 E. coli로부터의 세포 내 지질 추출" 한국화학공학회 51 (51): 482-486, 2013

2 Ellens, H., "pH-induced Destabilization of Phosphatidylethanolamine-containing Liposomes: Role of Bilayer Contact" 27 : 1532-1538, 1984

3 Struck, D. K., "Use of Resonance Energy Transfer to Monitor Membrane Fusion" 14 : 4093-4099, 1981

4 Schram, V., "Topology of Gel-phase Domains and Lipid Mixing Properties in Phase-separated Twocomponent Phosphatidylcholine Bilayers" 71 : 1811-1822, 1996

5 Ingolia, T. D., "The Role of Calcium in Fusion of Artificial Vesicles" 253 : 3821-3829, 1978

6 Huang, P., "The Potential for Phospholipase D as a New Therapeutic Target" 11 : 707-716, 2007

7 Park, J.-W., "Temperature Effect on Nanometer-scale Physical Properties of Mixed Phospholipid Monolayers" 62 : 157-161, 2008

8 Bentz, J., "Temperature Dependence of Divalent Cation Induced Fusion of Phosphatidylserine Liposomes: Evaluation of the Kinetic Rate Constants" 24 : 1064-1072, 1985

9 Gibson, G. A., "Phospholipid Vesicle Fusion Monitored by Fluorescence Energy Transfer" 88 : 135-140, 1979

10 Exton, J. H., "Phospholipase D: Structure, Regulation and Function" 144 : 1-94, 2002

11 Webb, L. M., "Phospholipase D Modulation by Ceramide in Senescence" 337 : 153-158, 2010

12 McDermott, M., "Phospholipase D" 82 : 225-253, 2004

13 Park, J.-W., "Phase Effect of Mixed-phospholipid Layer on Phospholipase D Reaction-induced-vesicle Rupture" 97 : 207-210, 2012

14 Tappia, P. S., "Oxidative Stress and Redox Regulation of Phospholipase D in Myocardial Disease" 41 : 349-361, 2006

15 Jin-Won Park, "Nanoliter Reactor Arrays for Antibiotic Study" 대한화학회 28 (28): 1709-1714, 2007

16 Vanderwerf, P., "Monitoring of Phospholipid Vesicle Fusion by Fluorescence Energy Transfer Between Membrane-bound Dye Labels" 596 : 302-314, 1980

17 New, R. R. C., "Liposomes: A Practical Approach" Academic Press 66-70, 1990

18 Brown, H. A., "Lipidomics and Bioactive Lipids: Lipids and Cell Signaling" Academic Press 58-100, 2007

19 Furt, F., "Importance of Lipid Metabolism for Intracellular and Mitochondrial Membrane Fusion/fission Processes" 41 : 1828-1836, 2009

20 Hoekstra, D., "Fusion of Phospholipid Vesicles Containing a Trypsin-sensitive Fluorogenic Substrate and Trypsin: a New Method to Study Membrane Fusion Activity in a Model System" 106 : 176-180, 1979

21 Uster, P. S., "Fusion Competence of Phosphatidylserine-containing Liposomes Quantitatively Measured by a Fluorescence Resonance Energy Transfer Assay" 209 : 385-395, 1981

22 Bongsoo Lee, "Enhancement of lipid productivity by ethyl methane sulfonate-mediated random mutagenesis and proteomic analysis in Chlamydomonas reinhardtii" 한국화학공학회 31 (31): 1036-1042, 2014

23 이길선, "Effect of Vesicle Curvature on Phospholipase D Reaction-Induced-Rupture" 대한화학회 34 (34): 3223-3226, 2013

24 Park, J.-W., "Effect of Phospholipid Bilayer Phase Asymmetry on Phospholipase D Reaction-induced Vesicle Rupture" 244 : 55-59, 2011

25 Scott, S. A., "Design of Isoform-selective Phospholipase D Inhibitors that Modulate Cancer Cell Invasiveness" 5 : 108-117, 2009

26 Park, J.-W., "Correlation Between Composition of the Outer Layer and Phase Asymmetry for Vesicles Ruptured by Phospholipase D" 246 : 399-405, 2013

27 Wilschut, J., "Ca2+-induced Fusion of Phospholipid Vesicles Monitored by Mixing of Aqueous Contents" 281 : 690-692, 1979

28 Lide, D. R., "CRC handbook of chemistry and physics: a readyreference book of chemical and physical data" CRC Press 220-246, 2005

29 Cheow, W. S., "Antibacterial Efficacy of Inhalable Antibiotic-Encapsulated Biodegradable Polymeric Nanoparticles against E. coli Biofilm Cells" 6 : 391-403, 2010

30 Owen, C. S., "A Membrane-bound Fluorescent Probe to Detect Phospholipid Vesicle-cell Fusion" 54 : 13-20, 1980

31 Keller, P. M., "A Fluorescence Enhancement Assay of Cell Fusion" 28 : 167-177, 1977