Roles of integrin β3 cytoplasmic tail in bidirectional signal transduction in a trans-dominant inhibition model

Abstract

We evaluated the roles of calpain cleavage-related mutations of the integrin β3 cytoplasmic tail in integrin αIIbβ3 bidirectional signaling using a trans-dominant inhibition model. Chimeric Tac-β3 proteins (i.e., Tac-β3, Tac-β3D741, Tac-β3D747, Tac-β3D754, Tac-β3D759, and Tac-β3DNITY) consisting of the extracellular and transmembrane domains of human IL-2 receptor (Tac) and the human integrin β3 cytoplasmic domain were stably expressed in the 123 CHO cells harboring human glycoprotein Ib-IX and wild-type integrin αIIbβ3. The different cells were assayed for stable adhesion and spreading on immobilized fibrinogen, and for binding soluble fibrinogen representing outside-in and inside-out signaling events, respectively. The chimeric protein Tac-β3 inhibited, and Tac-β3DNITY partially attenuated stable adhesion and spreading. Tac-β3, Tac-β3D759, Tac-β3DNITY, and Tac-β3D754, but not Tac-β3D747 or Tac-β3D741, impaired the soluble fibrinogen binding. Results indicated that the bidirectional signaling was significantly inhibited by Tac-β3 and Tac-β3DNITY, albeit to a much lesser extent. Moreover, only inside-out signaling was impaired in the 123/Tac-β3D759 and 123/Tac-β3D754 cells in contrast to an intact bidirectional signaling in the 123/Tac-β3D747 and 123/Tac-β3D741 cells. In conclusion, the calpain cleavage of integrin β3 resulted in the regulatory effects on signaling by interrupting its interaction with cytoplasmic proteins rather than altering its conformation, and may thus regulate platelet function.

Keywords

integrin β3 / signal transduction / trans-dominant inhibition model

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Acknowledgements

We thank all the laboratory members for helpful discussion. This work was supported by grants from National Natural Science Foundation of China (No. 81270594), National Basic Research Program of China (No. 2012CB518000, No. 2013CB966800) and National Natural Science Foundation of China (No. 81070414). Jiansong Huang is a recipient of a fellowship from The China Postdoctoral Science Foundation (No. 2013M531185).

Compliance with ethics guidelines

Jiansong Huang, Yulan Zhou, Xiaoyu Su, Yuanjing Lyu, Lanlan Tao, Xiaofeng Shi, Ping Liu, Zhangbiao Long, Zheng Ruan, Bing Xiao, Wenda Xi, Quansheng Zhou, Jianhua Mao, and Xiaodong Xi declare that they have no conflicts of interest. This article does not contain any studies with human or animal subjects performed by any of the authors.

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2016 Higher Education Press and Springer-Verlag Berlin Heidelberg