Franklin R A, Atherfold P A, Robinson P J, Bonner D
Department of Microbiology and Immunology, Brody School of Medicine at East Carolina University, Brody Building, Greenville, NC 27858, USA.
Cell Signal. 2001 Jan;13(1):65-9. doi: 10.1016/s0898-6568(00)00142-x.
Lysates from the Jurkat T lymphocyte cell line were immunoblotted with anti-Pyk2, and two major forms of Pyk2 were identified. When lysates from the p56(Lck) negative (J.CaM1/Rep3) and CD45 negative Jurkat cell line derivatives were immunoblotted with anti-Pyk2, only the lower mobility form of Pyk2 was predominant. Transfection of J.CaM1 cells with p56(Lck) restored expression of the multiple forms of Pyk2. Using RT-PCR, we found that both species of the alternatively spliced mRNA for Pyk2 were present in all of the lines regardless of their ability to express CD45 or p56(Lck) protein. When p56(Lck) immunoprecipitates were immunoblotted with anti-Pyk2, only the higher mobility form of Pyk2 immunoprecipitated with p56(Lck). These data demonstrate that certain members of the Src family of kinases interact preferentially with the different isoforms of Pyk2 and may have a role in the regulation of the Pyk2 protein in lymphocytes.
用抗Pyk2抗体对Jurkat T淋巴细胞系的裂解物进行免疫印迹分析,鉴定出Pyk2的两种主要形式。当用抗Pyk2抗体对p56(Lck)阴性(J.CaM1/Rep3)和CD45阴性的Jurkat细胞系衍生物的裂解物进行免疫印迹分析时,只有迁移率较低的Pyk2形式占主导。用p56(Lck)转染J.CaM1细胞可恢复Pyk2多种形式的表达。使用逆转录聚合酶链反应(RT-PCR),我们发现Pyk2的两种选择性剪接mRNA在所有细胞系中均存在,无论它们表达CD45或p56(Lck)蛋白的能力如何。当用抗Pyk2抗体对p56(Lck)免疫沉淀产物进行免疫印迹分析时,只有迁移率较高的Pyk2形式与p56(Lck)一起被免疫沉淀。这些数据表明,Src激酶家族的某些成员优先与Pyk2的不同异构体相互作用,可能在淋巴细胞中Pyk2蛋白的调节中发挥作用。