Effect of polylysine on transformations and permeability of negative vesicular membranes.
Small (40-60 nm in diameter) and large (300-350 nm) negative vesicles were complexed with a cationic polypeptide, poly-L-lysine (PL). Laser microelectrophoresis experiments showed that in small vesicles rendered anionic with the addition of cardiolipin (CL(2-)), only the CL(2-) in the outer leaflet is involved in the complexation with PL. Calorimetric and other data demonstrate that the binding of PL to the membrane surface causes domains ("rafts") of CL(2-) to form in the outer leaflet, and it is these domains that electrostatically bind the polymer. The kinetics of transmembrane permeation of doxorubicin (Dox, a fluorescent anti-tumor drug) was monitored with and without PL binding to the outer surface of the vesicles. It was found that PL mediates the permeation of Dox into the vesicle interior. In the absence of PL, the Dox molecule (possessing an amino group of pK(a)=8.6) binds to the anionic vesicles in the protonated form and, consequently, suffers an impaired mobility through the membrane. On the other hand, when the PL covers the vesicle surface, Dox passes though the membrane with greater ease. The effects of salt and polyanion on the stability of PL-vesicle complexes and the PL-mediated Dox permeation are also discussed.
Yaroslavov
A A
AA
School of Chemistry, M V Lomonosov Moscow State University, Leninskie Gory, Moscow 119899, Russian Federation. yaroslav@genebee.msu.su
Kuchenkova
O Ye
OY
Okuneva
I B
IB
Melik-Nubarov
N S
NS
Kozlova
N O
NO
Lobyshev
V I
VI
Menger
F M
FM
Kabanov
V A
VA
D015060:1,2-Dipalmitoylphosphatidylcholine; D000327:Adsorption; D000838:Anions; D000970:Antineoplastic Agents / Q000737:chemistry; D002308:Cardiolipins / Q000737:chemistry*; D003217:Conductometry; D004317:Doxorubicin / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008563:Membrane Lipids / Q000737:chemistry*; D010539:Permeability / Q000187:drug effects; D010713:Phosphatidylcholines / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry* / Q000494:pharmacology; D022161:Transport Vesicles / Q000737:chemistry / Q000187:drug effects
D000838:Anions; D000970:Antineoplastic Agents; D002308:Cardiolipins; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D011107:Polylysine; D015060:1,2-Dipalmitoylphosphatidylcholine; D004317:Doxorubicin