Chollet A, Bonnefoy J Y, Odermatt N
Glaxo Institute for Molecular Biology SA., Geneva, Switzerland.
J Immunol Methods. 1990 Mar 9;127(2):179-85. doi: 10.1016/0022-1759(90)90067-6.
Selective biotinylation of human interleukin-1 beta was achieved by reaction of an interleukin-1 beta mutant protein containing a surface-accessible cysteine with maleimido-biotin. A defined interleukin-1 beta derivative biotinylated at a single site was obtained. This compound retained full affinity for both the interleukin-1 receptor and streptavidin or avidin; however, its biological activity was significantly reduced in proliferative assays. Biotinylated interleukin-1 beta mutant protein bound to fluorescein-labelled avidin was used in flow cytometric analysis of interleukin-1 receptors. Internalization of the complex between biotinylated interleukin-1 beta mutant protein and streptavidin in a murine thymoma cell line was demonstrated.
通过使含有可及表面半胱氨酸的白细胞介素-1β突变蛋白与马来酰亚胺-生物素反应，实现了人白细胞介素-1β的选择性生物素化。获得了在单个位点生物素化的明确的白细胞介素-1β衍生物。该化合物对白细胞介素-1受体以及链霉亲和素或抗生物素蛋白均保持完全亲和力；然而，在增殖试验中其生物学活性显著降低。生物素化的白细胞介素-1β突变蛋白与荧光素标记的抗生物素蛋白结合，用于白细胞介素-1受体的流式细胞术分析。证明了生物素化的白细胞介素-1β突变蛋白与链霉亲和素之间的复合物在鼠胸腺瘤细胞系中的内化。
