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PMID: 11296243 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The ricinosomes of senescing plant tissue bud from the endoplasmic reticulum.

Schmid M, Simpson DJ, Sarioglu H, Lottspeich F, Gietl C

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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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2001-04-24
Epub
2001-00-10
Pages
5353-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC33213
Subset
IM
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