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EGY3: homologue of S2P protease located in chloroplasts.
Plant Biology ( IF 4.2 ) Pub Date : 2019-12-30 , DOI: 10.1111/plb.13087
M Adamiec 1 , L Misztal 1 , A Kasprowicz-Maluśki 2 , R Luciński 1
Affiliation  

  • The EGY3 protein is a homologue of site‐2 proteases, which are intramembrane zinc metalloproteases. EGY3 itself lacks proteolytic activity due to the absence of a zinc‐binding motif. Plentiful evidence indicates that such intramembrane ‘pseudoproteases’ play significant roles in many diverse processes occurring within the cell. However, the physiological functions of EGY3, as well as its subcellular localization, remain unknown.
  • The subcellular localization of EGY3 protein was investigated using Arabidopsis thaliana protoplasts transformed with EGY3‐GFP fusion protein, and immunoblot experiments using the total leaf protein extract, as well as highly purified chloroplasts and fractions of stroma, envelope and thylakoid membrane proteins. The physiological role of EGY3 was studied using two A. thaliana mutant lines devoid of EGY3 protein. Chlorophyll a fluorescence measurement was performed and the egy3 mutant sensitivity to photoinhibition was investigated. Additionally, the abundance of thylakoid membrane complexes was established using blue native gel electrophoresis.
  • We present experimental evidence for thylakoid membrane localization of the EGY3 protein.
  • We show that egy3 mutants display increased value of the non‐photochemical quenching parameter and significantly slower recovery rate after photoinhibitory treatment. This was associated with a decrease in the level of proteases involved in photosystem II recovery, Deg1 and FtsH2/8.


中文翻译:

EGY3:位于叶绿体中的S2P蛋白酶的同源物。

  • EGY3蛋白是Site-2蛋白酶的同系物,它们是膜内锌金属蛋白酶。由于缺乏锌结合基序,EGY3本身缺乏蛋白水解活性。大量证据表明,这种膜内“伪蛋白酶”在细胞内发生的许多不同过程中起重要作用。但是,EGY3的生理功能及其亚细胞定位仍然未知。
  • 使用转化为EGY3-GFP融合蛋白的拟南芥原生质体研究EGY3蛋白的亚细胞定位,并使用总叶蛋白提取物以及高度纯化的叶绿体和基质,包膜和类囊体膜蛋白的级分进行免疫印迹实验。使用两个不含EGY3蛋白的拟南芥突变株研究了EGY3的生理作用。叶绿素a荧光测量和egy3突变体对光抑制的敏感性进行了研究。另外,使用蓝色天然凝胶电泳建立了丰富的类囊体膜复合物。
  • 我们目前的EGY3蛋白类囊体膜定位的实验证据。
  • 我们显示,egy3突变体显示出非光化学猝灭参数的值增加,并且光抑制处理后的回收率显着降低。这与光系统II恢复所涉及的蛋白酶Deg1和FtsH2 / 8的水平降低有关。
更新日期:2019-12-30
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