Reduction of Plant-Specific Arabinogalactan-Type<i>O</i>-Glycosylation by Treating Tobacco Plants with Ferrous Chelator 2,2′-Dipyridyl

Moriguchi, Ryo; Matsuoka, Chiaya; Suyama, Akira; Matsuoka, Ken

doi:10.1271/bbb.100884

Cited by 12 publications

(9 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These data are consistent with reports on expression of recombinant EPO in Physcomitrella patens and a mucin-tandem repeat containing protein in N. benthamiana (29, 48). Future research directions will focus on strategies to prevent hydroxyproline formation by using inhibitors for prolyl 4-hydroxylases or specific knockdown/knock-out of the corresponding enzymes (32, 49, 50). …”

Section: Discussionmentioning

confidence: 99%

Engineering of Sialylated Mucin-type O-Glycosylation in Plants

Castilho

Neumann

Daskalova

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Plants lack the machinery for mucin-type O-glycosylation.Results: Transient expression of the mammalian O-glycosylation pathway in Nicotiana benthamiana resulted in the formation of sialylated mucin-type O-glycans on recombinant erythropoietin.Conclusion: Therapeutic proteins with engineered N- and O-glycosylation can be produced in plants.Significance: Plants are attractive hosts for the production of glycosylated recombinant proteins with defined glycan structures.

show abstract

Section: Discussionmentioning

confidence: 99%

Engineering of Sialylated Mucin-type O-Glycosylation in Plants

Castilho

Neumann

Daskalova

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…When needed, inhibitors of protein trafficking brefeldin A (BFA: Roche; final concentration 10 μg/ml) or wortmannin (Sigma–Aldrich; final concentration 10 μM), were added to the protoplast incubation medium 1 h after transfection and maintained at the same concentration throughout the incubation. To inhibit prolyl hydroxylation, transfected protoplasts were incubated in the presence of 0.25 mM 2,2′-dipyridyl (dipyr), a chemical that inhibits in vivo activity of prolyl-4-hydroxylase (Moriguchi et al, 2011). …”

Section: Methodsmentioning

confidence: 99%

Protein body formation in the endoplasmic reticulum as an evolution of storage protein sorting to vacuoles: insights from maize Î³-zein

et al. 2014

View full text Add to dashboard Cite

The albumin and globulin seed storage proteins present in all plants accumulate in storage vacuoles. Prolamins, which are the major proteins in cereal seeds and are present only there, instead accumulate within the endoplasmic reticulum (ER) lumen as very large insoluble polymers termed protein bodies. Inter-chain disulfide bonds play a major role in polymerization and insolubility of many prolamins. The N-terminal domain of the maize prolamin 27 kD γ-zein is able to promote protein body formation when fused to other proteins and contains seven cysteine residues involved in inter-chain bonds. We show that progressive substitution of these amino acids with serine residues in full length γ-zein leads to similarly progressive increase in solubility and availability to traffic from the ER along the secretory pathway. Total substitution results in very efficient secretion, whereas the presence of a single cysteine is sufficient to promote partial sorting to the vacuole via a wortmannin-sensitive pathway, similar to the traffic pathway of vacuolar storage proteins. We propose that the mechanism leading to accumulation of prolamins in the ER is a further evolutionary step of the one responsible for accumulation in storage vacuoles.

show abstract

“…Besides inactivating P4Hs expression, another approach would be to block hydroxylation of Pro residues using inhibitors. Moriguchi et al reported that the ferrous chelator 2,2'-dipyridyl, a potent inhibitor of P4Hs, reduced arabinogalactosylation of endogenous proteins in tobacco seedlings (125). Such a chemical inhibition strategy could be quite useful for transient expression but is less suitable for the stable expression of O-glycosylated recombinant proteins (94).…”

Section: O-glycosylation Differences Of O-glycosylation Between Humanmentioning

confidence: 99%

Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation

2016

View full text Add to dashboard Cite

Background and Purpose: Recombinant DNA technology has allowed expression of different heterologous proteins in many host systems, ranging from prokaryotic to eukaryotic organisms. Therapeutic properties of recombinant proteins are very often affected by the composition and heterogeneity of their glycans. Conventional expression systems for recombinant pharmaceuticals typically do not address this problem and result with products that contain a mixture of glycoforms that are neither identical to human glycans nor optimized for enhanced biological activity. Over the last decade plants have been developed as production platforms for recombinant proteins with pharmaceutical or industrial applications. Namely, plant expression systems contain very small differences in the post-translational modifications, mainly glycosylation, which can partly be overpowered by glycoengineering, whose goal is production of recombinant proteins with highly homogenous glycosylation that closely resembles the native system. This review attempts to present current accomplishments in the production of plant-derived glycoconjugates with humanized N-and O-glycans. Materials and Methods: Main goal of N-glycoengineering is to reduce or eliminate plant-specific N-glycans, and at the same time to introduce mammalian-specific N-glycans through the several approaches. The easiest way is to change intracellular targeting of plant-made recombinant proteins and to ensure their retention in the ER; next approach is to eliminate the addition of plant-specific glycans; while the final step is engineering the plant glycosylation pathway to introduce mammalian glycotransferases into plants with generation of biantennary and multi-antennary structures on complex N-glycans. Due to significant differences in O-glycosylation between humans and plants, different approaches to engineering of O-glycosylation have been taken. Besides having their typical O-glycoslyation on Hyp-residues, plants in general miss the machinery for production of mammalian-type O-glycosylation. Attempts have been made to mimic mammalian O-glycosylation in plants, specifically the mucin-type addition of GalNAc residues. Result: Efficient generation of bisected tetraantennary complex N-glycans without typical plant glycoepitopes on human erythropoietin (hEPO) and human transferrin (hTF) was obtained in Nicotiana benthamiana plants, thus demonstrating generation of recombinant proteins with human-type N-glycosylation at great uniformity. As for the O-glycosylation, attempts to produce mucin-type O-GalNAc and disialylated core 1 Olinked glycan structures on hEPO in N. benthamiana transgenic plants proved to be successful. Moreover, although small amounts of Hyp residues were found on recombinant EPO, no plant-specific O-glycans were de-IntroductIon R ecombinant DNA technology has allowed expression of different heterologous proteins in many host systems, ranging from prokaryotic to eukaryotic organisms. Recombinant microbial systems (mainly bacteria Escherichia coli) are in use ...

show abstract

Reduction of Plant-Specific Arabinogalactan-TypeO-Glycosylation by Treating Tobacco Plants with Ferrous Chelator 2,2′-Dipyridyl

Cited by 12 publications

References 16 publications

Engineering of Sialylated Mucin-type O-Glycosylation in Plants

Engineering of Sialylated Mucin-type O-Glycosylation in Plants

Protein body formation in the endoplasmic reticulum as an evolution of storage protein sorting to vacuoles: insights from maize Î³-zein

Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation

Contact Info

Product

Resources

About