Co‐expression of the protease furin in Nicotiana benthamiana leads to efficient processing of latent transforming growth factor‐β1 into a biologically active protein

分类： 生物学 >> 植物学 >> 植物生物化学、植物生物物理学 提交时间： 2016-05-04

Ruud H. P. Wilbers Lotte B. Westerhof Debbie R. van Raaij Marloes van Adrichem Andreas D. Prakasa Jose L. Lozano-Torres Jaap Bakker Geert Smant Arjen Schots

摘要： Transforming growth factor beta (TGF-) is a signalling molecule that plays a key role in developmental and immunological processes in mammals. Three TGF- isoforms exist in humans, and each isoform has unique therapeutic potential. Plants offer a platform for the production of recombinant proteins, which is cheap and easy to scale up and has a low risk of contamination with human pathogens. TGF-3 has been produced in plants before using a chloroplast expression system. However, this strategy requires chemical refolding to obtain a biologically active protein. In this study, we investigated the possibility to transiently express active human TGF-1 in Nicotiana benthamiana plants. We successfully expressed mature TGF-1 in the absence of the latency-associated peptide (LAP) using different strategies, but the obtained proteins were inactive. Upon expression of LAP-TGF-1, we were able to show that processing of the latent complex by a furin-like protease does not occur in planta. The use of a chitinase signal peptide enhanced the expression and secretion of LAP-TGF-1, and co-expression of human furin enabled the proteolytic processing of latent TGF-1. Engineering the plant post-translational machinery by co-expressing human furin also enhanced the accumulation of biologically active TGF-1. This engineering step is quite remarkable, as furin requires multiple processing steps and correct localization within the secretory pathway to become active. Our data demonstrate that plants can be a suitable platform for the production of complex proteins that rely on specific proteolytic processing.

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