PXD008538 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Three unrelated protease inhibitors enhance accumulation of pharmaceutical recombinant proteins in N. benthamiana |
Description | Agroinfiltrated Nicotiana benthamiana is a flexible and scalable recombinant protein (RP) production platform, but its great potential is hampered by plant proteases that degrade RPs. Here, we tested 29 candidate protease inhibitors (PIs) in agroinfiltrated N. benthamiana leaves for enhancing accumulation of three unrelated RPs: glycoenzyme α-Galactosidase, glycohormone erythropoietin (EPO) and IgG antibody VRC01. Of the previously described RP accumulation enhancing PIs, we found the cystatin SlCYS8 to be effective. We identified three additional new, unrelated PIs that enhanced RP accumulation: N. benthamiana NbPR4, NbPot1 and human HsTIMP, which are described to inhibit cysteine, serine and metalloproteases, respectively. Remarkably, accumulation of three unrelated RPs is enhanced by each PI, suggesting the mechanism of RP degradation may contain universal elements. Inhibitory motifs of HsTIMP and SlCYS8 are required to enhance RP accumulation, suggesting their target proteases may degrade RPs. Different PIs additively enhance RP accumulation, but the effect of each PI is dose-dependent. Activity-based Protein Profiling (ABPP) revealed that the activity profiles of Papain-like Cys proteases (PLCPs), Ser hydrolases (SHs) or Vacuolar Processing Enzymes (VPEs) in leaves are unaffected by the new PIs, whereas SlCYS8 specifically only suppresses PLCP activity. Quantitative leaf proteome data indicate that the three new PIs affect agroinfiltrated tissues similarly and that they all increase plant immunity. Our data indicate that RPs are degraded stepwise by a redundant, dynamic network of plant proteases. NbPR4, NbPot1 and HsTIMP can be used to identify and control new plant proteases and to enhance RP accumulation in industrial molecular farming. |
HostingRepository | PRIDE |
AnnounceDate | 2018-05-17 |
AnnouncementXML | Submission_2018-05-17_08:38:56.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Farnusch Kaschani |
SpeciesList | scientific name: Nicotiana benthamiana; NCBI TaxID: 4100; scientific name: Agrobacterium fabrum str. C58; NCBI TaxID: 176299; |
ModificationList | monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue |
Instrument | LTQ Orbitrap Elite |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2017-12-21 23:57:54 | ID requested | |
⏵ 1 | 2018-05-17 08:38:58 | announced | |
Publication List
Grosse-Holz F, Madeira L, Zahid MA, Songer M, Kourelis J, Fesenko M, Ninck S, Kaschani F, Kaiser M, van der Hoorn RAL, Three unrelated protease inhibitors enhance accumulation of pharmaceutical recombinant proteins in Nicotiana benthamiana. Plant Biotechnol J, 16(10):1797-1810(2018) [pubmed] |
Keyword List
curator keyword: Biological |
submitter keyword: Agrobacterium tumefaciens, biopharmaceutical production, transient expression, Activity Based Protein Profiling (ABPP), inhibitor mutant, metalloprotease |
Contact List
Farnusch Kaschani |
contact affiliation | Analytics Core Facility Essen (ACE), Chemische Biologie, Universität Duisburg-Essen, ZMB, Germany |
contact email | farnusch.kaschani@uni-due.de |
lab head | |
Farnusch Kaschani |
contact affiliation | University Duisburg-Essen |
contact email | farnusch.kaschani@uni-due.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD008538
- Label: PRIDE project
- Name: Three unrelated protease inhibitors enhance accumulation of pharmaceutical recombinant proteins in N. benthamiana