Identification and reconstitution of the rubber biosynthetic machinery on rubber particles from Hevea brasiliensis

Satoshi Yamashita, Haruhiko Yamaguchi, Toshiyuki Waki, Yuichi Aoki, Makie Mizuno, Fumihiro Yanbe, Tomoki Ishii, Ayuta Funaki, Yuzuru Tozawa, Yukino Miyagi-Inoue, Kazuhisa Fushihara, Toru Nakayama, Seiji Takahashi

研究成果: Article査読

71 被引用数 (Scopus)

抄録

Natural rubber (NR) is stored in latex as rubber particles (RPs), rubber molecules surrounded by a lipid monolayer. Rubber transferase (RTase), the enzyme responsible for NR biosynthesis, is believed to be a member of the cis-prenyltransferase (cPT) family. However, none of the recombinant cPTs have shown RTase activity independently. We show that HRT1, a cPT from Heveabrasiliensis, exhibits distinct RTase activity in vitro only when it is introduced on detergentwashed HeveaRPs (WRPs) by a cell-free translation-coupled system. Using this system, a heterologous cPT from Lactucasativa also exhibited RTase activity, indicating proper introduction of cPT on RP is the key to reconstitute active RTase. RP proteomics and interaction network analyses revealed the formation of the protein complex consisting of HRT1, rubber elongation factor (REF) and HRT1-REF BRIDGING PROTEIN. The RTase activity enhancement observed for the complex assembled on WRPs indicates the HRT1-containing complex functions as the NR biosynthetic machinery.

本文言語English
論文番号e19022
ジャーナルeLife
5
OCTOBER2016
DOI
出版ステータスPublished - 2016 10 28

ASJC Scopus subject areas

  • 神経科学(全般)
  • 免疫学および微生物学(全般)
  • 生化学、遺伝学、分子生物学(全般)

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