Gregorio-Jauregui Karla M, Carrizalez-Alvarez Susana A, Rivera-Salinas Jorge E, Saade Hened, Martinez José L, López Raúl G, Segura Elda P, Ilyina Anna
Grupo de Nanobiociencia de la Facultad de Ciencias Químicas, Universidad Autónoma de Coahuila, Boulevard V. Carranza y José Cárdenas Valdés, 25280, Saltillo, Coahuila, Mexico.
Appl Biochem Biotechnol. 2014 Apr;172(8):3721-35. doi: 10.1007/s12010-014-0801-x. Epub 2014 Feb 25.
The interaction between influenza virus hemagglutinins and host cell with terminal sialic acid linked receptors, SA-α-2,6-Gal for human strains is important to obtain insights into this infectious disease. Sambucus nigra lectin has high affinity for SA-α-2,6-Gal receptors. The goals of this work were: to extract the SA-α-2,6-Gal receptors from porcine airways; to perform receptors immobilization and study their storage stability; and to determine some parameters of interaction between the receptor and S. nigra lectin. The receptor isolation was monitored by means of bound sialic acid (BSAc) detection. A major band of protein at 66.7 kDa was clearly visible in SDS-PAGE assay. Eighty-one percent of isolated glycoproteins were immobilized on magnetic nanoparticles. The kinetics of BSAc storage stability at 4 °C was approximated as the first order reaction with kinetic constant and half-life estimated as 0.062 day(-1) and 11.2 days, respectively. The dissociation constant (K d) calculated from Scatchard's plot was 2.47 × 10(-7) M, and the receptor concentration was equal to 7.92 × 10(-5) M. Procedure for N-SA-α-2,6-Gal -receptors extraction based on their affinity to S. nigra lectin with magnetic nanoparticles, and their immobilization in active form, was not described previously, and may have wide application in designing biosensors or virus removal from areas or contaminated samples.
流感病毒血凝素与宿主细胞带有末端唾液酸连接受体（人株为SA-α-2,6-Gal）之间的相互作用，对于深入了解这种传染病至关重要。黑接骨木凝集素对SA-α-2,6-Gal受体具有高亲和力。本研究的目的是：从猪气道中提取SA-α-2,6-Gal受体；进行受体固定并研究其储存稳定性；以及确定受体与黑接骨木凝集素之间相互作用的一些参数。通过结合唾液酸（BSAc）检测来监测受体的分离。在SDS-PAGE分析中，66.7 kDa处可见一条主要蛋白带。81%的分离糖蛋白固定在磁性纳米颗粒上。4℃下BSAc储存稳定性的动力学近似为一级反应，动力学常数和半衰期分别估计为0.062天⁻¹和11.2天。根据Scatchard图计算的解离常数（Kd）为2.47×10⁻⁷ M，受体浓度等于7.92×10⁻⁵ M。基于其对带有磁性纳米颗粒的黑接骨木凝集素的亲和力提取N-SA-α-2,6-Gal受体及其以活性形式固定的方法，此前未见报道，可能在设计生物传感器或从区域或受污染样品中去除病毒方面有广泛应用。
