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Co-expression of the protease furin in Nicotiana benthamiana leads to efficient processing of latent transforming growth factor-b1 into a biologically active protein
- Source :
- Plant Biotechnology Journal, 14(8), 1695-1704, Plant Biotechnology Journal, Plant Biotechnology Journal 14 (2016) 8
- Publication Year :
- 2016
-
Abstract
- Summary Transforming growth factor beta (TGF‐β) is a signalling molecule that plays a key role in developmental and immunological processes in mammals. Three TGF‐β 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‐β3 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‐β1 in Nicotiana benthamiana plants. We successfully expressed mature TGF‐β1 in the absence of the latency‐associated peptide (LAP) using different strategies, but the obtained proteins were inactive. Upon expression of LAP‐TGF‐β1, 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‐β1, and co‐expression of human furin enabled the proteolytic processing of latent TGF‐β1. Engineering the plant post‐translational machinery by co‐expressing human furin also enhanced the accumulation of biologically active TGF‐β1. 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.
- Subjects :
- 0106 biological sciences
0301 basic medicine
medicine.medical_treatment
Nicotiana benthamiana
Plant Science
Immunoglobulin alpha-Chains
01 natural sciences
Protein Refolding
codon optimization
Furin
Research Articles
biology
transforming growth factor β1
food and beverages
Plants, Genetically Modified
PE&RC
Biochemistry
furin
Biotechnology
Research Article
Signal peptide
Recombinant Fusion Proteins
Protein Sorting Signals
Cell Line
Transforming Growth Factor beta1
03 medical and health sciences
Tobacco
medicine
Animals
Humans
Secretion
signal peptide
Laboratorium voor Nematologie
Secretory pathway
Protease
fungi
Epithelial Cells
Transforming growth factor beta
biology.organism_classification
Plant Leaves
030104 developmental biology
Mink
proteolytic processing
biology.protein
EPS
Laboratory of Nematology
Agronomy and Crop Science
010606 plant biology & botany
Transforming growth factor
Subjects
Details
- Language :
- English
- ISSN :
- 14677644
- Database :
- OpenAIRE
- Journal :
- Plant Biotechnology Journal, 14(8), 1695-1704, Plant Biotechnology Journal, Plant Biotechnology Journal 14 (2016) 8
- Accession number :
- edsair.doi.dedup.....75e57ba40ea675c7659db23a7330be8b