The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity

R.H.P. Wilbers, L.B. Westerhof, L.J. Reuter, A. Castilho, D.R. van Raaij, D. Nguyen, J.L. Lozano Torres, G. Smant, C.H. Hokke, J. Bakker, A. Schots

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)

Abstract

Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. IL-22 is an unusual cytokine as it does not act directly on immune cells. Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. Expression levels of IL-22 were up to 5.4 µg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ¿XT/FT plants a strong reduction in Lewis A was observed. Also, core a1,6-fucoylation did not improve the biological activity of IL-22. The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22.
Original languageEnglish
Pages (from-to)670-681
JournalPlant Biotechnology Journal
Volume14
Issue number2
DOIs
Publication statusPublished - 2016

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