Okamoto Takashi, Higuchi Kanako, Shinkawa Takashi, Isobe Toshiaki, Lörz Horst, Koshiba Tomokazu, Kranz Erhard
Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa 1-1, Hachioji, Tokyo, 192-0397 Japan.
Plant Cell Physiol. 2004 Oct;45(10):1406-12. doi: 10.1093/pcp/pch161.
In most flowering plants, the female gametophyte develops in an ovule deeply embedded in the ovary. Through double fertilization, the egg cell fuses with the sperm cell, resulting in a zygote, which develops into the embryo. In the present study, we analyzed egg cell lysates by polyacrylamide gel electrophoresis and subsequent mass spectrometry-based proteomics technology, and identified major protein components expressed in the egg cell. The identified proteins included three cytosolic enzymes of the glycolytic pathway, glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase and triosephosphate isomerase, two mitochondrial proteins, the ATP synthase beta-subunit and an adenine nucleotide transporter, and annexin p35. In addition, expression levels of these proteins in the egg cell were compared with those in the early embryo, the central cell and the suspension cell. Annexin p35 was highly expressed only in the egg cell, and glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase and the adenine nucleotide transporter were expressed at higher levels in egg cells than in central and cultured cells. These results indicate that annexin p35 in the egg cell and zygote is involved in the exocytosis of cell wall materials, which is induced by a fertilization-triggered increase in cytosolic Ca2+ levels, and that the egg cell is rich in an enzyme subset for the energy metabolism.
在大多数开花植物中,雌配子体在深深嵌入子房的胚珠中发育。通过双受精,卵细胞与精细胞融合,形成合子,合子进而发育成胚胎。在本研究中,我们通过聚丙烯酰胺凝胶电泳和随后基于质谱的蛋白质组学技术分析了卵细胞裂解物,并鉴定了卵细胞中表达的主要蛋白质成分。鉴定出的蛋白质包括糖酵解途径的三种胞质酶,即甘油醛-3-磷酸脱氢酶、3-磷酸甘油酸激酶和磷酸丙糖异构酶,两种线粒体蛋白,ATP合酶β亚基和腺嘌呤核苷酸转运体,以及膜联蛋白p35。此外,还比较了这些蛋白质在卵细胞与早期胚胎、中央细胞和悬浮细胞中的表达水平。膜联蛋白p35仅在卵细胞中高表达,甘油醛-3-磷酸脱氢酶、3-磷酸甘油酸激酶和腺嘌呤核苷酸转运体在卵细胞中的表达水平高于中央细胞和培养细胞。这些结果表明,卵细胞和受精卵中的膜联蛋白p35参与了细胞壁物质的胞吐作用,这是由受精引发的胞质Ca2+水平升高所诱导的,并且卵细胞富含能量代谢的一组酶。