PXD008538

PXD008538 is an original dataset announced via ProteomeXchange.

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

Revision	Datetime	Status	ChangeLog Entry
0	2017-12-21 23:57:54	ID requested
⏵ 1	2018-05-17 08:38:58	announced

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]

curator keyword: Biological

submitter keyword: Agrobacterium tumefaciens, biopharmaceutical production, transient expression, Activity Based Protein Profiling (ABPP), inhibitor mutant, metalloprotease

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

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PRIDE project URI

• PRIDE
  1. PXD008538
     1. Label: PRIDE project
     2. Name: Three unrelated protease inhibitors enhance accumulation of pharmaceutical recombinant proteins in N. benthamiana