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Coagulation & its Disorders
Structural and cellular mechanisms of peptidyl-prolyl isomerase Pin1-mediated enhancement of Tissue Factor gene expression, protein half-life, and pro-coagulant activity
Kondababu Kurakula,1,2,* Duco S. Koenis,1,* Mark A. Herzik Jr,3,4 Yanyun Liu,3 John W. Craft Jr,3 Pieter B. van Loenen,1 Mariska Vos,1 M. Khang Tran,1 Henri H. Versteeg,5 Marie-José T.H. Goumans,2 Wolfram Ruf,6,7
Carlie J.M. de Vries1,# and Mehmet Şen3,#
1Department of Medical Biochemistry, Amsterdam Cardiovascular Sciences, Academic Medical Center, University of Amsterdam, the Netherlands; 2Department of Cell and Chemical Biology, Leiden University Medical Center, the Netherlands; 3Department of Biology and Biochemistry, Chemical Biology Interdisciplinary Program, University of Houston, TX, USA; 4Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, USA; 5The Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, the Netherlands; 6Center for Thrombosis and Hemostasis, University Medical Center, Mainz, Germany and 7Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA, USA
ABSTRACT
Tissue Factor is a cell-surface glycoprotein expressed in various cells of the vasculature and is the principal regulator of the blood coag- ulation cascade and hemostasis. Notably, aberrant expression of Tissue Factor is associated with cardiovascular pathologies such as ath- erosclerosis and thrombosis. Here, we sought to identify factors that reg- ulate Tissue Factor gene expression and activity. Tissue Factor gene expression is regulated by various transcription factors, including activat- ing protein-1 and nuclear factor-κ B. The peptidyl-prolyl isomerase Pin1 is known to modulate the activity of these two transcription factors, and we now show that Pin1 augments Tissue Factor gene expression in both vascular smooth muscle cells and activated endothelial cells via activat- ing protein-1 and nuclear factor-κ B signaling. Furthermore, the cytoplas- mic domain of Tissue Factor contains a well-conserved phospho-Ser258- Pro259 amino-acid motif recognized by Pin1. Using co-immunoprecipi- tation and solution nuclear magnetic resonance spectroscopy, we show that the WW-domain of Pin1 directly binds the cytoplasmic domain of Tissue Factor. This interaction occurs via the phospho-Ser258-Pro259 sequence in the Tissue Factor cytoplasmic domain and results in increased protein half-life and pro-coagulant activity. Taken together, our results establish Pin1 as an upstream regulator of Tissue Factor-mediated coagulation, thereby opening up new avenues for research into the use of specific Pin1 inhibitors for the treatment of diseases characterized by pathological coagulation, such as thrombosis and atherosclerosis.
Introduction
Tissue Factor (TF), an integral cell-surface glycoprotein, is the initiator of the blood coagulation cascade and a key regulator of hemostasis.1,2 Aberrant expression of TF plays a crucial role in several coagulation-driven pathologies, such as thrombosis, ath- erosclerosis, and acute coronary syndromes,2-4 but also in endotoxemia, angiogenesis, and cancer.5-8 Many vascular cells express TF constitutively, including smooth muscle cells (SMCs), pericytes, and adventitial fibroblasts, while TF expression is unde- tectable in vascular endothelial cells (ECs).1 However, in both ECs and SMCs, the expression and activity of TF can be enhanced by pro-inflammatory signaling mole-
Ferrata Storti Foundation
Haematologica 2018 Volume 103(6):1073-1082
*KK and DSK contributed equally to this work. #CJMdV and MS contributed equally to this work.
Correspondence:
c.j.devries@amc.nl or msen@uh.edu
Received: October 23, 2017. Accepted: March 1, 2018. Pre-published: March 15, 2018.
doi:10.3324/haematol.2017.183087
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/103/6/1073
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