{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:50:38Z","timestamp":1791420638862,"version":"4.3.3"},"reference-count":64,"publisher":"eLife Sciences Publications, Ltd","license":[{"start":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T00:00:00Z","timestamp":1477612800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T00:00:00Z","timestamp":1477612800000},"content-version":"am","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T00:00:00Z","timestamp":1477612800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:p>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 detergent-washed 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.<\/jats:p>","DOI":"10.7554\/elife.19022","type":"journal-article","created":{"date-parts":[[2016,10,28]],"date-time":"2016-10-28T08:00:14Z","timestamp":1477641614000},"source":"Crossref","is-referenced-by-count":154,"title":["Identification and reconstitution of the rubber biosynthetic machinery on rubber particles from Hevea brasiliensis"],"prefix":"10.7554","volume":"5","author":[{"given":"Satoshi","family":"Yamashita","sequence":"first","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haruhiko","family":"Yamaguchi","sequence":"additional","affiliation":[{"name":"Sumitomo Rubber Industries, Ltd, Kobe, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshiyuki","family":"Waki","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuichi","family":"Aoki","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makie","family":"Mizuno","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fumihiro","family":"Yanbe","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomoki","family":"Ishii","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayuta","family":"Funaki","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuzuru","family":"Tozawa","sequence":"additional","affiliation":[{"name":"Graduate School of Science and Engineering, Saitama University, Saitama, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukino","family":"Miyagi-Inoue","sequence":"additional","affiliation":[{"name":"Sumitomo Rubber Industries, Ltd, Kobe, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuhisa","family":"Fushihara","sequence":"additional","affiliation":[{"name":"Sumitomo Rubber Industries, Ltd, Kobe, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toru","family":"Nakayama","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2288-4340","authenticated-orcid":true,"given":"Seiji","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Tohoku University, Sendai, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4374","published-online":{"date-parts":[[2016,10,28]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1111\/tpj.12063","article-title":"The tomato cis-prenyltransferase gene family","volume":"73","author":"Akhtar","year":"2013","journal-title":"The Plant Journal"},{"key":"bib2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.plantsci.2014.05.003","article-title":"Identification of laticifer-specific genes and their promoter regions from a natural rubber producing plant Hevea brasiliensis","volume":"225","author":"Aoki","year":"2014","journal-title":"Plant Science"},{"key":"bib3","doi-asserted-by":"publisher","first-page":"593","DOI":"10.5511\/plantbiotechnology.14.1015a","article-title":"Transcriptional responses of laticifer-specific genes to phytohormones in a suspension-cultured cell line derived from petioles of Hevea brasiliensis","volume":"31","author":"Aoki","year":"2014","journal-title":"Plant Biotechnology"},{"key":"bib4","first-page":"476","volume-title":"Methods Enzymol","author":"Archer","year":"1969"},{"key":"bib5","doi-asserted-by":"publisher","first-page":"4671","DOI":"10.1046\/j.1432-1033.2003.03863.x","article-title":"Molecular cloning, expression and characterization of cDNA encoding cis-prenyltransferases from Hevea brasiliensis. 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