Dubois T, Oudinet J P, Russo-Marie F, Rothhut B
Laboratoire de Signalisation, Inflammation et Transformation Cellulaire, INSERM U.332, Institut Cochin de Génétique Moléculaire (ICGM), Université René Descartes, Paris, France.
Biochem J. 1995 Aug 15;310 ( Pt 1)(Pt 1):243-8. doi: 10.1042/bj3100243.
In order to understand how signal transduction occurs during T cell activation, it is necessary to identify the key regulatory molecules whose function is influenced by phosphorylation. Annexins II (A-II) and V (A-V) belong to a large family of Ca(2+)-dependent phospholipid-binding proteins. Among many putative functions, annexins may be involved in signal transduction during cellular proliferation and differentiation. In the present study we show that A-II is phosphorylated in vivo in the Jurkat human T cell line. Indeed, A-II is phosphorylated after stimulation by phorbol myristate acetate and on serine residues after T cell antigen receptor (TcR) stimulation. In cytosol from Jurkat cells, A-II is phosphorylated only by Ca2+/phospholipid-stimulated kinases such as Ca(2+)-dependent protein kinases C (cPKCs). A-V inhibits the phosphorylation of A-II and other substrates of cPKCs and has no effect on kinases activated only by phospholipids. In conclusion, A-II is phosphorylated both in vitro and in vivo in Jurkat cells, and may play a role as a substrate during signal transduction in lymphocytes via the TcR through the PKC pathway. On the other hand, A-V could act as a potent modulator of cPKCs in Jurkat cells.
为了了解T细胞激活过程中信号转导是如何发生的,有必要鉴定其功能受磷酸化影响的关键调节分子。膜联蛋白II(A-II)和V(A-V)属于一个依赖Ca(2+)的磷脂结合蛋白大家族。在众多假定的功能中,膜联蛋白可能参与细胞增殖和分化过程中的信号转导。在本研究中,我们发现A-II在Jurkat人T细胞系中发生体内磷酸化。事实上,佛波酯肉豆蔻酸酯刺激后以及T细胞抗原受体(TcR)刺激后,A-II在丝氨酸残基上发生磷酸化。在Jurkat细胞的胞质溶胶中,A-II仅被Ca2+/磷脂刺激的激酶如依赖Ca(2+)的蛋白激酶C(cPKCs)磷酸化。A-V抑制A-II和cPKCs其他底物的磷酸化,并且对仅由磷脂激活的激酶没有影响。总之,A-II在Jurkat细胞中体外和体内均发生磷酸化,并且可能在淋巴细胞通过TcR经PKC途径的信号转导过程中作为底物发挥作用。另一方面,A-V可能在Jurkat细胞中作为cPKCs的有效调节剂发挥作用。