Chakrabarti A, Podder S K
Department of Biochemistry, Indian Institute of Science, Bangalore.
Biochim Biophys Acta. 1990 May 9;1024(1):103-10. doi: 10.1016/0005-2736(90)90213-8.
Two types of phospholipid vesicles capable of mutual recognition have been tailor-made to serve as a model system for the study of carbohydrate-mediated cellular adhesion. One of the vesicles contained a fatty acid conjugate of a galactose specific lectin (lectin vesicle) and the other an asialoganglioside with a reactive terminal galactose residue (galactose vesicle). The kinetics of aggregation of these two types of vesicles was followed by monitoring time-dependent change in turbidity. A 10-100-fold enhancement in the forward rate constant (kf ranging from 7.1 x 10(5) to 4.5 x 10(7) M-1.s-1 at 27 degrees C) was observed when compared with that for the lectin-galactose system in solution (kf being 4.5 x 10(5) M-1.s-1), reported in the literature. A study of the influence of vesicle size on the rate of aggregation showed that enhancement depended on the curvature of the galactose vesicle rather than the density of asialoganglioside suggesting a possible diffusion in the plane of the membrane. The ratio, kf/kd is found to be approx. 10(10) M-1 indicating that the formation of multiple bonds plays a role for stable adhesion.
两种能够相互识别的磷脂囊泡已被特制出来,用作研究碳水化合物介导的细胞黏附的模型系统。其中一种囊泡含有半乳糖特异性凝集素的脂肪酸结合物(凝集素囊泡),另一种含有带有反应性末端半乳糖残基的去唾液酸神经节苷脂(半乳糖囊泡)。通过监测随时间变化的浊度来跟踪这两种囊泡的聚集动力学。与文献报道的溶液中凝集素 - 半乳糖系统(27℃时正向速率常数kf为4.5×10⁵ M⁻¹·s⁻¹)相比,观察到正向速率常数有10 - 100倍的提高(27℃时kf范围为7.1×10⁵至4.5×10⁷ M⁻¹·s⁻¹)。对囊泡大小对聚集速率影响的研究表明,这种提高取决于半乳糖囊泡的曲率,而不是去唾液酸神经节苷脂的密度,这表明可能在膜平面内发生扩散。发现kf/kd的比值约为10¹⁰ M⁻¹,表明多键的形成对稳定黏附有作用。