Subcellular proteome profiles of different latex fractions revealed washed solutions from rubber particles contain crucial enzymes for natural rubber biosynthesis.
Subcellular proteome profiles of different latex fractions revealed washed solutions from rubber particles contain crucial enzymes for natural rubber biosynthesis.
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DOI：
10.1016/j.jprot.2018.05.002
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发表时间：
2018-06
影响因子：
3.3
通讯作者：
Dan Wang;Yong Sun;Lili Chang;Zheng Tong;Quanliang Xie;Xiang Jin;Liping Zhu;P. He;Hongbin Li;Xuchu Wang
中科院分区：
文献类型：
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作者：
Dan Wang;Yong Sun;Lili Chang;Zheng Tong;Quanliang Xie;Xiang Jin;Liping Zhu;P. He;Hongbin Li;Xuchu Wang
橡胶颗粒（RP）是橡胶树橡胶树（hevea brasiliensis）天然橡胶合成和储存的特异性细胞器。NRB由RP膜定位蛋白处理，传统上通过反复洗涤纯化。然而，我们注意到RP丢弃的洗涤液（WS）中有许多蛋白质。在此，我们用2-DE比较了WS、c -血清（CS）和RP的蛋白质组谱，并通过质谱分析从WS中鉴定出233个丰富的蛋白质。2-DE凝胶上的许多斑点被鉴定为不同的蛋白质种类。我们进一步对CS、WS和RP进行了霰弹枪分析，分别鉴定出1837、1799和1020个独特蛋白。加上2-DE，我们最终从WS中鉴定出1825个蛋白，其中246个是WS特异性蛋白。这些ws特异性蛋白在Gene Ontology中进行了注释，表明最丰富的途径是有机物代谢过程、蛋白质降解过程、初级代谢过程和能量代谢过程。蛋白-蛋白相互作用分析表明，这些ws特异性蛋白主要参与核糖体代谢、蛋白酶体系统、空泡蛋白分选和内吞作用。无标记和Western blotting结果显示，许多ws特异性蛋白和蛋白复合物对NRB的起始起着至关重要的作用。这些发现不仅加深了我们对WS蛋白质组的认识，也为RP膜蛋白在NRB中的作用提供了新的证据。天然橡胶储存在橡胶树的橡胶颗粒中。橡胶颗粒的传统纯化是通过反复洗涤，但从洗涤液（WS）中鉴定出许多蛋白质。我们首次获得了1825个WS蛋白的可视化蛋白质组图谱，其中包括246个WS特异性蛋白。这些WS蛋白几乎包含所有聚异戊二烯起始酶，可能在橡胶生物合成中发挥重要作用。
Rubber particle (RP) is a specific organelle for natural rubber biosynthesis (NRB) and storage in rubber treeHevea brasiliensis. NRB is processed by RP membrane-localized proteins, which were traditionally purified by repeated washing. However, we noticed many proteins in the discarded washing solutions (WS) from RP. Here, we compared the proteome profiles of WS, C-serum (CS) and RP by 2-DE, and identified 233 abundant proteins from WS by mass spectrometry. Many spots on 2-DE gels were identified as different protein species. We further performed shotgun analysis of CS, WS and RP and identified 1837, 1799 and 1020 unique proteins, respectively. Together with 2-DE, we finally identified 1825 proteins from WS, 246 were WS-specific. These WS-specific proteins were annotated in Gene Ontology, indicating most abundant pathways are organic substance metabolic process, protein degradation, primary metabolic process, and energy metabolism. Protein-protein interaction analysis revealed these WS-specific proteins are mainly involved in ribosomal metabolism, proteasome system, vacuolar protein sorting and endocytosis. Label free and Western blotting revealed many WS-specific proteins and protein complexes are crucial for NRB initiation. These findings not only deepen our understanding of WS proteome, but also provide new evidences on the roles of RP membrane proteins in NRB.SignificanceNatural rubber is stored in rubber particle from the rubber tree. Rubber particles were traditionally purified by repeated washing, but many proteins were identified from the washing solutions (WS). We obtained the first visualization proteome profiles with 1825 proteins from WS, including 246 WS-specific ones. These WS proteins contain almost all enzymes for polyisoprene initiation and may play important roles in rubber biosynthesis.