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目的:研究接头蛋白(adaptorprotein)Bam32在B细胞抗原受体(Bcellantigenreceptor,BCR)信号转导级联反应中的作用。方法:以Bam32全序列为诱饵,应用酵母双杂交技术筛选能与Bam32相互作用的蛋白分子,并用293T细胞共转染和免疫共沉淀法加以证实。结果:应用酵母双杂交技术筛选出能与Bam32相互作用的蛋白分子,其中1个出现强阳性反应的克隆编码酪氨酸激酶Lyn。用293T细胞共转染和特异性免疫共沉淀法证实,Bam32可在哺乳动物细胞中与Lyn共沉淀。应用抗磷酸化酪氨酸抗体检测显示,Bam32与Lyn相互作用可导致Bam32磷酸化。结论:Bam32可同Lyn相互作用,导致Bam32磷酸化,这在激活下游产物的级联反应中可能起重要作用。
AIM: To investigate the role of adapter protein Bam32 in Bcellantigen receptor (BCR) signal transduction cascade. Methods: The Bam32 full sequence was used as bait. The yeast two - hybrid system was used to screen the protein molecules that could interact with Bam32. The results were confirmed by co - transfection of 293T cells and co - immunoprecipitation. Results: The yeast two-hybrid technique was used to screen out the protein molecules that could interact with Bam32. One of the strong positive reaction clones encoded tyrosine kinase Lyn. Cotransfection with 293T cells and specific co-immunoprecipitation confirmed that Bam32 co-precipitated with Lyn in mammalian cells. Detection using anti-phospho-tyrosine antibodies showed that Bam32 interacts with Lyn and leads to Bam32 phosphorylation. CONCLUSION: Bam32 interacts with Lyn, resulting in phosphorylation of Bam32, which may play an important role in the cascade of downstream products.