DOI: 10.1016/s0014-5793(03)00916-5. A transient tobacco expression system coupled to MALDI‐TOF‐MS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants

A transient tobacco expression system coupled to MALDI‐TOF‐MS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants

10.1016/s0014-5793(03)00916-5

Crossref journal-article

Wiley

FEBS Letters (311)

Abstract

Tobacco‐based transient expression was employed to elucidate the impact of differential targeting to subcellular compartments on activity and quality of gastric lipase as a model for the production of recombinant glycoproteins in plants. Overall N‐linked glycan structures of recombinant lipase were analyzed and for the first time sugar structures of its four individual N‐glycosylation sites were determined in situ by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) on a trypsin digest without isolation or deglycosylation of the peptides. Three glycosylation sites contain both complex‐type N‐glycans and high‐mannose‐type structures, the fourth is exclusively linked to high‐mannose glycans. Although the overall pattern of glycan structures is influenced by the targeting, our results show that the type of glycans found linked to a given Asn residue is largely influenced by the physico‐chemical environment of the site. The transient tobacco system combined with MALDI‐TOF‐MS appears to be a useful tool for the evaluation of glycoprotein production in plants.

Bibliography

Mokrzycki-Issartel, N., Bouchon, B., Farrer, S., Berland, P., Laparra, H., Madelmont, J.-C., & Theisen, M. (2003). A transient tobacco expression system coupled to MALDIâTOFâMS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants. FEBS Letters, 552(2â3), 170â176. Portico.

Authors 7

Nathalie Mokrzycki-Issartel (first)
Bernadette Bouchon (additional)
Sibille Farrer (additional)
Patricia Berland (additional)
Hélène Laparra (additional)
Jean-Claude Madelmont (additional)
Manfred Theisen (additional)

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Dates

Type	When
Created	21 years, 11 months ago (Sept. 11, 2003, 9:10 p.m.)
Deposited	1 year, 11 months ago (Sept. 16, 2023, 10:55 p.m.)
Indexed	1 year, 10 months ago (Oct. 9, 2023, 2:15 p.m.)
Issued	22 years ago (Aug. 27, 2003)
Published	22 years ago (Aug. 27, 2003)
Published Online	22 years ago (Aug. 27, 2003)
Published Print	21 years, 11 months ago (Sept. 25, 2003)

Funders 0

None

BibTeX

@article{Mokrzycki_Issartel_2003, title={A transient tobacco expression system coupled to MALDI‐TOF‐MS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants}, volume={552}, ISSN={1873-3468}, url={http://dx.doi.org/10.1016/s0014-5793(03)00916-5}, DOI={10.1016/s0014-5793(03)00916-5}, number={2–3}, journal={FEBS Letters}, publisher={Wiley}, author={Mokrzycki-Issartel, Nathalie and Bouchon, Bernadette and Farrer, Sibille and Berland, Patricia and Laparra, Hélène and Madelmont, Jean-Claude and Theisen, Manfred}, year={2003}, month=aug, pages={170–176} }

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