Enhancing Tissue Factor Production: The Role of N-Glycosylation and ERAD Pathway Modulation

Yi-Shi Liu; Yue Dou; Xiaoman Zhou; Zijie Li; Nakanishi Hideki

doi:10.2174/0109298665364078250519065417

Enhancing Tissue Factor Production: The Role of N-Glycosylation and ERAD Pathway Modulation

Protein Pept Lett. 2025 May 27. doi: 10.2174/0109298665364078250519065417. Online ahead of print.

Authors

Yi-Shi Liu¹, Yue Dou¹, Xiaoman Zhou¹, Zijie Li¹, Nakanishi Hideki¹

Affiliation

¹ Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu214122, China.

PMID: 40444633
DOI: 10.2174/0109298665364078250519065417

Abstract

Background: Tissue Factor (TF) is a crucial transmembrane glycoprotein that triggers blood coagulation upon vascular or tissue injury by binding to plasma factors VII and VIIa. In recent years, the demand for TF has rapidly increased due to its pivotal role in preoperative coagulation tests. However, large-scale production of TF remains challenging despite successful recombinant expression, as incorrect post-translational modifications adversely affect TF activity.

Objective: This study aims to investigate the role of post-translational modifications, specifically N-glycosylation, in TF activity and stability. Additionally, it explores strategies to enhance TF production by reducing its degradation through genetic modulation.

Methods: We compared TF activity derived from human cells and E. coli to assess the impact of post-translational modifications. Furthermore, we examined the effect of N-glycosylation on TF function. To address TF degradation, we knocked out the HRD1 gene, a key component of the endoplasmic- reticulum-associated degradation (ERAD) pathway, and evaluated its impact on TF stability and activity.

Results: TF produced in human cells exhibited higher activity than TF expressed in E. coli, emphasizing the importance of post-translational modifications. Specifically, N-glycosylation was found to influence TF activity and stability significantly. Additionally, we observed that knocking out the HRD1 gene effectively reduced TF degradation without compromising its activity.

Conclusion: Our findings underscore the crucial role of N-glycosylation in TF function and stability. Moreover, the modulation of the ERAD pathway through HRD1 knockout presents a promising approach for enhancing TF production. These insights could contribute to the large-scale manufacturing of functionally active TF for clinical and research applications.

Keywords: Blood coagulation; ERAD; N-glycosylation; Tissue factor; coagulation test; preoperative test..