Lactose-installed poly(ethylene glycol)-poly(D,L-lactide) block copolymer micelles exhibit fast-rate binding and high affinity toward a protein bed simulating a cell surface. A surface plasmon resonance study

被引:82
作者
Jule, E
Nagasaki, Y
Kataoka, K
机构
[1] Univ Tokyo, Dept Mat Sci & Engn, Grad Sch Engn, Bunkyo Ku, Tokyo 1138656, Japan
[2] Tokyo Univ Sci, Dept Mat Sci, Noda, Chiba 2788510, Japan
关键词
D O I
10.1021/bc025598+
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Lactose molecules were installed on the surface of poly(ethylene glycol)-poly(D,L-lactide) (PEG-PLA) block copolymer micelles in the scope of seeking specific recognition by cell surface receptors at hepatic sites. This, in turn, is expected to result in the formation of a complex displaying prolonged retention times and thus enhanced cellular internalization by receptor-mediated endocytosis. The so-obtained particles based on a block copolymer of molecular weight 9400 g/mol (4900/4500 g/mol for the PEG and PLA blocks, respectively) were found to have an average hydrodynamic diameter of 31.8 nm, as measured by dynamic light scattering. Further, the particle size distribution (mu(2)/(Gamma) over bar (2)) was found to be lower than 0.08. Lactose-PEG-PLA micelles (Lac-micelles) were then injected over a gold surface containing Ricinus communis agglutinin lectins simulating the aforementioned glycoreceptors, and their interaction was studied by surface plasmon resonance. Then, a kinetic evaluation was carried out, by fitting the observed data mathematically. It appears that Lac-micelles bind in a multivalent manner to the lectin protein bed, which logically results in low dissociation constants. Micelles bearing a ligand density of 80% (Lac-micelles 80%: 80 lactose molecules per 100 copolymer chains) exhibit fast association phases (k(a1) = 3.2 x 10(4) M(-1) s(-1)), but also extremely slow dissociation phases (k(d1) = 1.3 x 10(-4) s(-1)). Recorded sensorgrams were fitted with a trivalent model, conveying a calculated equilibrium dissociation constant (K(D1) = k(d1)/k(a1)) of about 4 nM. The importance of cooperative binding was also assessed, by preparing Lac-micelles bearing different ligand densities, and by discussing the influence of the latter on kinetic constants. Interestingly enough, whereas Lac-micelles 80% bind in a trivalent manner to the protein bed, Lac-micelles 20% are still capable of forming bivalent complexes with the same protein bed (KD1 = 1360 nM). Therefore, despite enhanced kinetic values brought about by a supplementary bond, lower ligand densities appear to be more effective on a molecular basis.
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页码:177 / 186
页数:10
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