Schmid M, Simpson D J, Sarioglu H, Lottspeich F, Gietl C
Lehrstuhl für Botanik, Biologikum-Weihenstephan, Technische Universität München, Am Hochanger 4, D-85350 Freising, Germany.
Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5353-8. doi: 10.1073/pnas.061038298. Epub 2001 Apr 10.
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.
蓖麻毒蛋白体(同义词,前体蛋白酶小泡)是一种新型细胞器,迄今为止仅在植物细胞中发现。电子显微镜研究表明,它在衰老组织中从内质网出芽形成。现在,对这种不同寻常起源的生化支持来自纯化细胞器的组成,该细胞器含有大量具有C末端KDEL基序的45 kDa半胱氨酸内肽酶前体以及内质网腔驻留蛋白BiP(结合蛋白)和蛋白质二硫键异构酶。蛋白质免疫印迹分析、肽测序和质谱分析表明KDEL保留在蛋白酶原形式中。分离的蓖麻毒蛋白体的酸化会导致蓖麻籽半胱氨酸内肽酶激活,伴随N末端前肽和C末端KDEL基序的切割。我们提出,蓖麻毒蛋白体在衰老过程中通过程序性细胞死亡积累,并通过酸性液泡释放质子而被激活。