Abstract
The ricinosome (synonym, precursor protease vesicle) is a novel organelle, found so far exclusively in plant cells. Electron microscopic studies suggest that it buds off from the endoplasmic reticulum in senescing tissues. Biochemical support for this unusual origin now comes from the composition of the purified organelle, which contains large amounts of a 45-kDa cysteine endoprotease precursor with a C-terminal KDEL motif and the endoplasmic reticulum lumen residents BiP (binding protein) and protein disulfide isomerase. Western blot analysis, peptide sequencing, and mass spectrometry demonstrate retention of KDEL in the protease proform. Acidification of isolated ricinosomes causes castor bean cysteine endopeptidase activation, with cleavage of the N-terminal propeptide and the C-terminal KDEL motif. We propose that ricinosomes accumulate during senescence by programmed cell death and are activated by release of protons from acidic vacuoles.
MeSH Terms
Amino Acid Sequence
Apoptosis
Blotting, Western
Carrier Proteins/analysis
Centrifugation, Density Gradient
Cyanogen Bromide/metabolism
Cysteine Endopeptidases/chemistry,metabolism
Cytoplasmic Vesicles/chemistry,enzymology,metabolism,ultrastructure
Endoplasmic Reticulum/chemistry,enzymology,metabolism,ultrastructure
Endoplasmic Reticulum Chaperone BiP
Enzyme Activation
Fluorescent Antibody Technique
Heat-Shock Proteins
Hydrogen-Ion Concentration
Mass Spectrometry
Microscopy, Electron
Molecular Chaperones/analysis
Molecular Sequence Data
Molecular Weight
Oligopeptides/metabolism
Peptide Fragments/chemistry,metabolism
Plants, Toxic
Protein Disulfide-Isomerases/metabolism
Protein Precursors/chemistry,metabolism
Protein Processing, Post-Translational
Protein Sorting Signals
Ricinus/cytology,growth & development,ultrastructure
Vacuoles/metabolism
Chemicals
Carrier Proteins
Endoplasmic Reticulum Chaperone BiP
Heat-Shock Proteins
Molecular Chaperones
Oligopeptides
Peptide Fragments
Protein Precursors
Protein Sorting Signals
lysyl-aspartyl-glutamyl-leucine
Cysteine Endopeptidases
Protein Disulfide-Isomerases
Cyanogen Bromide
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Schmid M
Lehrstuhl für Botanik, Biologikum-Weihenstephan, Technische Universität München, Am Hochanger 4, D-85350 Freising, Germany.
Simpson D J
Sarioglu H
Lottspeich F
Gietl C
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