A cysteine endopeptidase with a C-terminal KDEL motif isolated from castor bean endosperm is a marker enzyme for the ricinosome, a putative lytic compartment.
A papain-type cysteine endopeptidase with a molecular mass of 35 kDa for the mature enzyme, was purified from germinating castor bean (Ricinus communis L.) endosperm by virtue of its capacity to process the glyoxysomal malate dehydrogenase precursor protein to the mature subunit in vitro (C. Gietl et al., 1997, Plant Physiol 113: 863-871). The cDNA clones from endosperm of germinating seedlings and from developing seeds were isolated and sequence analysis revealed that a very similar or identical peptidase is synthesised in both tissues. Sequencing established a presequence for co-translational targeting into the endoplasmic reticulum, an N-terminal propeptide and a C-terminal KDEL motif for the castor bean cysteine endopeptidase precursor. The 45-kDa pro-enzyme stably present in isolated organelles was enzymatically active. Immunocytochemistry with antibodies raised against the purified cysteine endopeptidase revealed highly specific labelling of ricinosomes, organelles which co-purify with glyoxysomes from germinating Ricinus endosperm. The cysteine endopeptidase from castor bean endosperm, which represents a senescing tissue, is homologous to cysteine endopeptidases from other senescing tissues such as the cotyledons of germinating mung bean (Vigna mungo) and vetch (Vicia sativa), the seed pods of maturing French bean (Phaseolus vulgaris) and the flowers of daylily (Hemerocallis sp.).
Lehrstuhl für Botanik, Technische Universität München, Germany.
M
M
Schmid
D
D
Simpson
F
F
Kalousek
C
C
Gietl
D001483:Base Sequence; D015415:Biomarkers; D002367:Ricinus communis / Q000166:cytology / Q000201:enzymology*; D002451:Cell Compartmentation; D002499:Centrifugation, Density Gradient; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D018076:DNA, Complementary; D018744:DNA, Plant; D004792:Enzyme Precursors / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides; D015388:Organelles / Q000201:enzymology* / Q000648:ultrastructure; D010947:Plants, Toxic; D021382:Protein Sorting Signals; D013395:Sucrose
D015415:Biomarkers; D018076:DNA, Complementary; D018744:DNA, Plant; D004792:Enzyme Precursors; D009842:Oligopeptides; D021382:Protein Sorting Signals; C073762:lysyl-aspartyl-glutamyl-leucine; D013395:Sucrose; D003546:Cysteine Endopeptidases