Open AccessJournal Article
Multiple functional proteins are produced by cleaving asn-gln bonds of a single precursor by vacuolar processing enzyme
TLDR
The results suggested that VPE was responsible for cleaving Asn-Gln bonds of a single precursor, PV100, to produce multiple seed proteins, and it is likely that the Asn -Gln stretches not only provide cleavage sites for VPE but also produce aminopeptidase-resistant proteins.Abstract:
Precursor-accumulating vesicles mediate transport of the precursors of seed proteins to protein storage vacuoles in maturing pumpkin seeds. We isolated the precursor-accumulating vesicles and characterized a 100-kDa component (PV100) of the vesicles. Isolated cDNA for PV100 encoded a 97,310-Da protein that was composed of a hydrophobic signal peptide and the following three domains: an 11-kDa Cys-rich domain with four CXXXC motifs, a 34-kDa Arg/Glu-rich domain composed of six homologous repeats, and a 50-kDa vicilin-like domain. Both immunocytochemistry and immunoblots with anti-PV100 antibodies showed that <10-kDa proteins and the 50-kDa vicilin-like protein were accumulated in the vacuoles. To identify the mature proteins derived from PV100, soluble proteins of the vacuoles were separated, and their molecular structures were determined. Mass spectrometry and peptide sequencing showed that two Cys-rich peptides, three Arg/Glu-rich peptides, and the vicilin-like protein were produced by cleaving Asn-Gln bonds of PV100 and that all of these proteins had a pyroglutamate at their NH2 termini. To clarify the cleavage mechanism, in vitro processing of PV100 was performed with purified vacuolar processing enzyme (VPE). Taken together, these results suggested that VPE was responsible for cleaving Asn-Gln bonds of a single precursor, PV100, to produce multiple seed proteins. It is likely that the Asn-Gln stretches not only provide cleavage sites for VPE but also produce aminopeptidase-resistant proteins. We also found that the Cys-rich peptide functions as a trypsin inhibitor. Our findings suggested that PV100 is converted into different functional proteins, such as a proteinase inhibitor and a storage protein, in the vacuoles of seed cells.read more
Citations
More filters
Journal ArticleDOI
Cysteine proteases of parasitic organisms
Mohammed Sajid,James H. McKerrow +1 more
TL;DR: In this paper, the authors highlight recent research on the Papain-like and asparaginyl-endopeptidase classes of cysteine proteases and re-examine them in light of the diversity uncovered within parasitic organisms.
Journal ArticleDOI
A plant vacuolar protease, VPE, mediates, virus-induced hypersensitive cell death
Noriyuki Hatsugai,Miwa Kuroyanagi,Kenji Yamada,Kenji Yamada,Tetsuo Meshi,Shinya Tsuda,Maki Kondo,Mikio Nishimura,Mikio Nishimura,Ikuko Hara-Nishimura +9 more
TL;DR: VPE deficiency prevented virus-induced hypersensitive cell death in tobacco plants and showed that plants have evolved a regulated cellular suicide strategy that, unlike PCD of animals, is mediated by VPE and the cellular vacuole.
Journal ArticleDOI
A cut above the rest: the regulatory function of plant proteases.
TL;DR: Recent findings for the major catalytic classes, i.e. the serine, cysteine, aspartic, and metalloproteases, are reviewed, emphasizing the regulatory function of representative enzymes.
Journal ArticleDOI
Proteomics of Medicago truncatula Seed Development Establishes the Time Frame of Diverse Metabolic Processes Related to Reserve Accumulation
TL;DR: A proteomic approach was utilized to investigate seed development in Medicago truncatula and revealed a differential accumulation of enzymes involved in methionine metabolism and proposed a role for these enzymes in the transition from a highly active to a quiescent state during seed development.
Journal ArticleDOI
Vacuolar processing enzyme: an executor of plant cell death.
TL;DR: Plants might have evolved a regulated cellular suicide strategy that, unlike animal apoptosis, is mediated by VPE and the vacuoles.
References
More filters
Journal ArticleDOI
Accumulation of β-conglycinin in soybean cotyledon through the formation of disulfide bonds between α'- and α-subunits.
Hiroyuki Wadahama,Kensuke Iwasaki,Motonori Matsusaki,Keito Nishizawa,Masao Ishimoto,Fumio Arisaka,Kyoko Takagi,Reiko Urade +7 more
TL;DR: The results suggest that disulfide bonds are formed between α′/α-sub units residing in different β-conglycinin hexamers, but the binding of P34 to α′- and α-subunits reduces the linkage between β-Conglycin in hexamer.
Journal ArticleDOI
The N-terminal ricin propeptide influences the fate of ricin A-chain in tobacco protoplasts.
Nicholas A. Jolliffe,Alessandra Di Cola,Catherine J. Marsden,J. Michael Lord,Aldo Ceriotti,Lorenzo Frigerio,Lynne M. Roberts +6 more
TL;DR: It is demonstrated that the N-terminal propeptide of proricin acts as a nonspecific spacer to ensure efficient ER import and glycosylation, and that when absent from the N terminus of ricin A-chain, the non-imported material remained tethered to the cytosolic face of the ER membrane.
Journal ArticleDOI
Elevated field atmospheric CO2 concentrations affect the characteristics of winter wheat (cv. Bologna) grains
Francesca Verrillo,Franz-Werner Badeck,Valeria Terzi,F. Rizza,Letizia Bernardo,Antimo Di Maro,Clara Fares,Alessandro Zaldei,Francesco Miglietta,Anna Moschella,Marcella Bracale,Candida Vannini +11 more
TL;DR: The results suggest that the expected increase in CO2 levels and their interactive effects with environmental variables may influence agronomic performance by increasing yield and negatively affecting quality.
Journal ArticleDOI
Isolation and biochemical characterization of two forms of RD21 from cotyledons of daikon radish (Raphanus sativus).
TL;DR: In this paper, an Arabidopsis cysteine protease with a granulin-like domain at the C-terminus was purified from the cotyledons of daikon radish (Raphanus sativus) and the expression of the enzyme was initiated by imbibition and continued at almost constant levels during germination.
Journal ArticleDOI
Peptide Fragments From Plant Vicilins Expressed in Escherichia Coli Exhibit Antimicrobial Activity In Vitro
TL;DR: Histidine-tagged versions of the putative antimicrobial peptides were expressed in Escherichia coli, purified, and demonstrated to have in vitro antimicrobial activity.