TheBiophysicalSociety of Japan General IncorporatedAssociation suggest that surface properties o uptake into a GUV, fPLGA-NPs are important for endoeytosis-like 3PTt40 Hirotaka Ariyama, Graduate Slrhooi qfStience and Tlechnology, Transportan permeation membranes using LUV suspension have been characteristics membranes remain unclear. 10 on membrane unitameltar vesic]e First, we investigated the dioleoylphosphatidylcholine(DOPC) containing the contrast, fiuorescence microscopy. DOPC-GUVsinduced

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... he rapid monotonous leakage ofca]cein from of the most During the calcein leakage, the size ofmost GUVs decreased a litt]e (less than S %), but in some GUVs their radius decreased greatly and some small hlghcontrast particles appeared on the GUV membranes. The fraction of leaked GUVincreased with time, and aiso with an increase in TP 10 concentration, Seeond, the effect of cholesterol on the TP 1O-induced leakage ofca]cein was investigated. O.2 pM TP 10 did not induce ca]cein ]eakage from DOPCIchol (6I4)-GUVs. On the basis of these data, we discuss the mechan{sm of the interaction ofTP 1O with DOPC-GUVs and DOPC/chol-GUVs. FJyx nt-s ;t 1o pl Dopc utoE ± u utv-hopmmtatte tlliBt[ljZ6suM Effects of Transportan-10 on Membrane Permeability and Structure of Single Giant Unilame]lar Vesicles of DOPC membranes Masahito Yamuzaki (lntegrated Bio.seience Section, S]iizuoka Uhiv.) 10 {TP 10) is one of the cel] penetrating peptides,Se far the of TP 10 into cells and the inteTaction of TP 10 with ]ipid investigated. However, the detai] and mechanism of the interaction and its pemieation through In this report, we have investigated thc effeet of TP perrneability of lipid membranes using the sing]e giant (GUV)method. interaction of TP 1O with sing]e GUVs composed of and PEG2K-DOPE (mo]ar ratio, 98:2) fluorescent dye, calcein. in a physiological buffer using phase The interaction of O.8 pM TP lO with sing]e the inside GUVs, but in some GUVs step-wise leakages were observed. 3PT141 F-BARIISUUrtY-AOf=-IL,-YiYopU7JVStAMM Real-time observation ef liposome tubu]ation by F-BAR Yohko Takiguchii, Toshiki Itoh2, Kingo Takiguchii (iGrad. Sch. Sci., Uhiv, Nbgaya, 2Grad. Sch. Med., Uhiv. Kobe) The Fer-CIP4 homology-BAR (F-BAR) domain has been identit'ied as a biological membrane-deforming module. The F-BAR domain has been Teported to transform lipid bilayer membranes into tubules. However, the precess of tubutatien still remains unknewn, Here we monitored the entire tubulatien process indueed by the F-BAR domains or by the fu1Mengths of four different F-BAR dornain proteins, PSTPIPI, FBP17, CIP4 and Pacsin2, using direet rea]time irnaging, and show that each F-BAR domain or protein induces tubules through a unique k{netics ftem mode] membranes. FBPr7 and CIP4 develop many prejections simultaneous]y throughout the surfhce of indiividual liposomes, whereas PSTPIP1 and Pacsin2 deve]op on]y a few projections from a narrow restricted part of the surface of individuat ]iposornes. The resu]ts provide striking cvidcnce that a nucleation process is involvedintheF-BAR-inducedtubulation,andindividualF-BARdomainshave a unique nucleation rate andror property even though essentialty they have the same crescent-shapcd structurc. The differences in process of tubulation induced with those F-BAR domain proteins may refiect their unique physiolegical retes, and function favorabty te buitd networks of varieus and robust mernbrane traencking processes observed in cells. pEGRrevv(7)FtNtvoJvt[as[iummnte"wrartcocaee Correlation between Membrane Composition and Mode of Division of PEG-grafted Giant Vesicles , Kensuke Kurihara?, Taro Teyota2'i, Masayuki lmai4, Tadashi (iOchanemizu Uhiversity, 21Vie Uhiversity of' 7bkvo, 3Reseat'ch Complex S,stems Bioiogy, 47bhoku Uhiversity, SKbnagau,a division. Here we prepared a new modet protocett inctuding S mel% of polyethylene g]yeol {PEG5000)-gTafted phospholipid/ a PEG-chain of the phosphetipid not only increased the tolerance ofGV to high ionic strength and high temperature but also affected the interaction between DNA and the GV membrane containing the cationic membTane mo]ecu]e V, In order to elucidate the morphological ehange of the GV membrane throughout the setil reproduction process, we stained the mernbrane by Texas Red-tagged phospho]ipid and observed thc dynamics in terms of a confoeal laser scanning fluorescence micrescope, Whereas the division mode of non PEG-grafted GV was a budding-type exc]us[vely, the birthing-type deformation occurred rnainly in the case of PEG-grafted GV. We specu]ated that the interaction between the ainplified DNA and the PEG-grafted inner surface ofthe membrane influences the division mode ofGV selrreproduction. In this symposium, we wi[] discuss the cfTbct of the ]ength and the content of PEG-grafted phospholipid on the mode ofGV division. 3PTt43 Takeuehi2 {dKanagawa Acadentv qf Science and 7lechnology, 2institute qf' industrial st'ience, 7)be University oj'7bb'o) Thisworkpresentsamethodo]ogythatrealizessize-controlled,solvent-freegiant liposome arrays with simple gent]e hydration technique. Gent]e hydratjon and electroforrnation have been the most common]y used methods to prepare giant liposomes, yet they have dienculty controlling the size distribution of the formed liposemes as well as their shape and ]amellarity, We thercfore set the goa] of developing an alternative methed that allows the formation ofuniform-size giant tiposomes with high repToducibility. Our system consists ofa dried lipid pattern on a substrate that is a micropatterned po]ymer thin film on an ITO glass slide. An electrospray deposition {ESD) method was applied to obtain the selective ]ipiddeposition at the polymer pattcrn where the ITO surface was exposed. With a simp]e hydration proeess of the dried lipid, we succeeded in formation ofgiant tiposomcs on top of the pattern. The diameter ofthe 1iposomes was sirnilar to the patterned size, achieying a narrow range of size distribution. We also report the applicability of thc system for vurious bio]ogical assays such as object encapsu]ation into the liposomes, buffer exchange during the assays, and reconstitution of membrane proteins into the ]iposome membranes. 3PT142 Yumi Ka"i SUgawara]'S Center for Uhiversity) Recentty 3PT144 REMDV:-=b-YiYXS6Mzatt"ntercfiutopreMBO-fi Study oil phase transition of coarse-grained lipid bitayer by REMD simulations Tetsuro Nagai, Yuko Okamoto (Grad. Sch. Sci., Nbgaya Uhiv.) A replica-exchange molecular dynamics simulation ofa lipid bilayer system was perfonned in order to study sol-gel phase transitions. The rep]ica-exchange molecuiar dynamies method enables one te enhance conformational samp]ing efficiency and to study the system at a wide teinperature range at once. We ernployed a coarse-grained mode] MARTINI. The results show abrupt ehanges in interna] energy, bilayer thickness, and area of bitayer around 296 K. A peak jn specifie head capacjty was also observed around this temperature. These suggested that the sol-gel phase transitions. The bilayer has two stateg in the gel state. One is a ti]ted ge] state and the other is an un-ti]ted ge] state, A pTevious work with MARTINI force filed reported only the un-titled ge] state. This indicates that con formational sampling cfFlciency is not trivia] even with coarsegrained modet, which has smeether eneTgy landscapes and reach ]enger time scales than atomistic modet. The un-tilted gel phase is a little thicker than the tilted ge] phase, whilc thc arca oflipid biluyer is quite similar. We will show more detail results in the presentation. 3PT145 ficaU;,vafiDMPCefisuU;,rsfiDHPCEMnt"nUrereza re\llopS'( ± EOX Dynamics of complicated phase behavior on the mixtures consisting ef long-chain phospho]ipids DMPC and short-chain phospholipids DHPC Ryota Kobayashi. Tetsuhiko Ohba (Department (tf Physics, Tbhoku Uhiver.siij,.) The binary phospho]ipid mixtures of long-chain DMPC and short-chain DHPC show a variety of comp]icated phase behavior, depending on the mixing ratio of two ]ipids, the tota] tipid conccntration in water, and temperature. At low we realized a model protocell using a giant vesic[e {GV) as a compartment, comprised of phospholipids, cationic synthesized mernbrane molecule (V) and an amphiphilic eata]yst (C). Encapsu]ated DNA in GV was ampljfied by pelyrnerase chain reaction, and then the GV grew and divided when the incorporated membrane-precursor was converted to V by the assistance of C. We a]so revealed that the attractive interaction between amplified DNA and the inner surface ofthe GV membrane acce]erated the GV -S165 -NII-Electronic