PXD030443-1

PXD030443 is an original dataset announced via ProteomeXchange.

Title	Unexpected arabinosylation after humanization of plant protein N-glycosylation
Description	As biopharmaceuticals, recombinant proteins have become indispensable tools in medicine. An increasing demand, not only in quantity but also in diversity, drives the constant development and improvement of production platforms. The N-glycosylation pattern on biopharmaceuticals plays important roles in activity, serum half-life and immunogenicity. Therefore, production platforms with tailored protein N-glycosylation are of great interest. Plant-based systems have already demonstrated their potential to produce pharmaceutically relevant recombinant proteins, although their N-glycan patterns differ from those in humans. Plants have shown great plasticity towards the manipulation of their glycosylation machinery, and some have already been glyco-engineered in order to avoid the attachment of plant-typical, putatively immunogenic sugar residues. This resulted in complex-type N-glycans with a core structure identical to the human one. Compared to humans, plants lack the ability to elongate these N-glycans with β1,4-linked galactoses and terminal sialic acids. However, these modifications, which require the activity of several mammalian enzymes, have already been achieved for Nicotiana benthamiana and the moss Physcomitrella. Here, we present the first step towards sialylation of recombinant glycoproteins in Physcomitrella, human β1,4-linked terminal N-glycan galactosylation, which was achieved by the introduction of a chimeric β1,4-galactosyltransferase (FTGT). This chimeric enzyme consists of the moss α1,4-fucosyltransferase transmembrane domain, fused to the catalytic domain of the human β1,4-galactosyltransferase. Stable FTGT expression led to the desired β1,4-galactosylation. However, additional pentoses of unknown identity were also observed. The nature of these pentoses was subsequently determined by Western blot and enzymatic digestion followed by mass spectrometric analysis and resulted in their identification as α-linked arabinoses. Since a pentosylation of β1,4-galactosylated N-glycans was reported earlier, e.g. on recombinant human erythropoietin produced in glyco-engineered Nicotiana tabacum, this phenomenon is of a more general importance for plant-based production platforms. Arabinoses, which are absent in humans, may prevent the full humanization of plant-derived products. Therefore, the identification of these pentoses as arabinoses is important as it creates the basis for their abolishment to ensure the production of safe biopharmaceuticals in plant-based systems.
HostingRepository	PRIDE
AnnounceDate	2024-10-08
AnnouncementXML	Submission_2024-10-08_11:38:08.545.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ralf Reski
SpeciesList	scientific name: Physcomitrella; NCBI TaxID: 3217;
ModificationList	iodoacetamide derivatized residue
Instrument	Q Exactive; 6520A Quadrupole Time-of-Flight LC/MS

Revision	Datetime	Status	ChangeLog Entry
0	2021-12-16 07:09:03	ID requested
⏵ 1	2024-10-08 11:38:09	announced

Bohlender LL, Parsons J, Hoernstein SNW, Bangert N, Rodr, í, guez-Jahnke F, Reski R, Decker EL, -Glycosylation. Front Bioeng Biotechnol, 10():838365(2022) [pubmed]

10.3389/fbioe.2022.838365;

submitter keyword: Physcomitrella (Physcomitrium patens), plant-made recombinant biopharmaceuticals,4-galactosylation,glyco-optimization, N-glycan humanization, glyco-engineering, plant-made pharmaceuticals, β1, N-glycan pentosylation

Ralf Reski
contact affiliation	Head, Chair Plant Biotechnology Faculty of Biology University of Freiburg Schaenzlestrasse 1 D-79104 Freiburg Germany
contact email	ralf.reski@biologie.uni-freiburg.de
lab head
Ralf Reski
contact affiliation	Faculty of Biology, University of Freiburg (Chair Plant Biotechnology), Schaenzlestr. 1, D-79104 Freiburg
contact email	ralf.reski@biologie.uni-freiburg.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/04/PXD030443

PRIDE project URI

• PRIDE
  1. PXD030443
     1. Label: PRIDE project
     2. Name: Unexpected arabinosylation after humanization of plant protein N-glycosylation