Wang Sai, Liu Jiahui, Wu Yanxia, You Yawen, He Jingyi, Zhang Jichuan, Zhang Liqun, Dong Yiyang
Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
Center of Advanced Elastomer Materials, College of Materials Science and Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
Anal Biochem. 2016 Apr 15;499:34-42. doi: 10.1016/j.ab.2016.01.015. Epub 2016 Feb 2.
Commercial natural rubber is traditionally supplied by Hevea brasiliensis, but now there is a big energy problem because of the limited resource and increasing demand. Intensive study of key rubber-related substances is urgently needed for further research of in vitro biosynthesis of natural rubber. Natural rubber is biosynthesized on the surface of rubber particles. A membrane protein called small rubber particle protein (SRPP) is a key protein associated closely with rubber biosynthesis; however, SRPP in different plants has been only qualitatively studied, and there are no quantitative reports so far. In this work, H. brasiliensis was chosen as a model plant. The microscopic distribution of SRPP on the rubber particles during the washing process was investigated by transmission electron microscopy-immunogold labeling. A label-free surface plasmon resonance (SPR) immunosensor was developed to quantify SRPP in H. brasiliensis for the first time. The immunosensor was then used to rapidly detect and analyze SRPP in dandelions and prickly lettuce latex samples. The label-free SPR immunosensor can be a desirable tool for rapid quantitation of the membrane protein SRPP, with excellent assay efficiency, high sensitivity, and high specificity. The method lays the foundation for further study of the functional relationship between SRPP and natural rubber content.
传统上，商业天然橡胶由巴西橡胶树供应，但由于资源有限和需求增加，现在存在一个重大的能源问题。为了进一步研究天然橡胶的体外生物合成，迫切需要对关键橡胶相关物质进行深入研究。天然橡胶在橡胶粒子表面生物合成。一种名为小橡胶粒子蛋白（SRPP）的膜蛋白是与橡胶生物合成密切相关的关键蛋白；然而，不同植物中的SRPP仅进行了定性研究，目前尚无定量报道。在这项工作中，选择巴西橡胶树作为模式植物。通过透射电子显微镜免疫金标记研究了洗涤过程中SRPP在橡胶粒子上的微观分布。首次开发了一种无标记表面等离子体共振（SPR）免疫传感器来定量巴西橡胶树中的SRPP。然后使用该免疫传感器快速检测和分析蒲公英和刺莴苣乳胶样品中的SRPP。这种无标记SPR免疫传感器可以成为快速定量膜蛋白SRPP的理想工具，具有出色的检测效率、高灵敏度和高特异性。该方法为进一步研究SRPP与天然橡胶含量之间的功能关系奠定了基础。
