Identification of aTaraxacum brevicorniculatumrubber elongation factor protein that is localized on rubber particles and promotes rubber biosynthesis

2015 ◽  
Vol 82 (4) ◽  
pp. 609-620 ◽  
Author(s):  
Natalie Laibach ◽  
Andrea Hillebrand ◽  
Richard M. Twyman ◽  
Dirk Prüfer ◽  
Christian Schulze Gronover
Keyword(s):  
Elongation Factor ◽  
Rubber Biosynthesis ◽  
Rubber Particles

scholarly journals Proteomic Landscape Has Revealed Small Rubber Particles Are Crucial Rubber Biosynthetic Machines for Ethylene-Stimulation in Natural Rubber Production

2019 ◽  
Vol 20 (20) ◽  
pp. 5082 ◽  
Author(s):  
Dan Wang ◽  
Quanliang Xie ◽  
Yong Sun ◽  
Zheng Tong ◽  
Lili Chang ◽  
...  
Keyword(s):  
Natural Rubber ◽  
Rubber Particle ◽  
Family Members ◽  
Elongation Factor ◽  
Absolute Quantification ◽  
Protein Accumulation ◽  
Rubber Biosynthesis ◽  
Rubber Particles ◽  
Related Proteins ◽  
Latex Production

Rubber particles are a specific organelle for natural rubber biosynthesis (NRB) and storage. Ethylene can significantly improve rubber latex production by increasing the generation of small rubber particles (SRPs), regulating protein accumulation, and activating many enzyme activities. We conducted a quantitative proteomics study of different SRPs upon ethylene stimulation by differential in-gel electrophoresis (DIGE) and using isobaric tags for relative and absolute quantification (iTRAQ) methods. In DIGE, 79 differentially accumulated proteins (DAPs) were determined as ethylene responsive proteins. Our results show that the abundance of many NRB-related proteins has been sharply induced upon ethylene stimulation. Among them, 23 proteins were identified as rubber elongation factor (REF) and small rubber particle protein (SRPP) family members, including 16 REF and 7 SRPP isoforms. Then, 138 unique phosphorylated peptides, containing 129 phosphorylated amino acids from the 64 REF/SRPP family members, were identified, and most serine and threonine were phosphorylated. Furthermore, we identified 226 DAPs from more than 2000 SRP proteins by iTRAQ. Integrative analysis revealed that almost all NRB-related proteins can be detected in SRPs, and many proteins are positively responsive to ethylene stimulation. These results indicate that ethylene may stimulate latex production by regulating the accumulation of some key proteins. The phosphorylation modification of REF and SRPP isoforms might be crucial for NRB, and SRP may act as a complex natural rubber biosynthetic machine.

scholarly journals Amino Acid Sequence of Rubber Elongation Factor Protein Associated with Rubber Particles in Hevea Latex

1989 ◽  
Vol 264 (31) ◽  
pp. 18618-18626 ◽  
Author(s):  
M S Dennis ◽  
W J Henzel ◽  
J Bell ◽  
W Kohr ◽  
D R Light
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Elongation Factor ◽  
Rubber Particles

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

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Satoshi Yamashita ◽  
Haruhiko Yamaguchi ◽  
Toshiyuki Waki ◽  
Yuichi Aoki ◽  
Makie Mizuno ◽  
...  
Keyword(s):  
Elongation Factor ◽  
Interaction Network ◽  
Coupled System ◽  
Lipid Monolayer ◽  
Network Analyses ◽  
Rubber Particles ◽  
Free Translation ◽  
A Cell ◽  
Biosynthetic Machinery

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.

scholarly journals Proteomic Landscape of the Mature Roots in a Rubber-Producing Grass Taraxacum Kok-saghyz

2019 ◽  
Vol 20 (10) ◽  
pp. 2596 ◽  
Author(s):  
Quanliang Xie ◽  
Guohua Ding ◽  
Liping Zhu ◽  
Li Yu ◽  
Boxuan Yuan ◽  
...  
Keyword(s):  
Natural Rubber ◽  
Large Scale ◽  
Rubber Particle ◽  
Elongation Factor ◽  
Regulation Mechanism ◽  
Cascade Process ◽  
Two Dimensional Gel Electrophoresis ◽  
Rubber Biosynthesis ◽  
Phenylpropanoid Biosynthesis ◽  
Related Proteins

The rubber grass Taraxacum kok-saghyz (TKS) contains large amounts of natural rubber (cis-1,4-polyisoprene) in its enlarged roots and it is an alternative crop source of natural rubber. Natural rubber biosynthesis (NRB) and storage in the mature roots of TKS is a cascade process involving many genes, proteins and their cofactors. The TKS genome has just been annotated and many NRB-related genes have been determined. However, there is limited knowledge about the protein regulation mechanism for NRB in TKS roots. We identified 371 protein species from the mature roots of TKS by combining two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Meanwhile, a large-scale shotgun analysis of proteins in TKS roots at the enlargement stage was performed, and 3545 individual proteins were determined. Subsequently, all identified proteins from 2-DE gel and shotgun MS in TKS roots were subject to gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses and most proteins were involved in carbon metabolic process with catalytic activity in membrane-bounded organelles, followed by proteins with binding ability, transportation and phenylpropanoid biosynthesis activities. Fifty-eight NRB-related proteins, including eight small rubber particle protein (SRPP) and two rubber elongation factor(REF) members, were identified from the TKS roots, and these proteins were involved in both mevalonate acid (MVA) and methylerythritol phosphate (MEP) pathways. To our best knowledge, it is the first high-resolution draft proteome map of the mature TKS roots. Our proteomics of TKS roots revealed both MVA and MEP pathways are important for NRB, and SRPP might be more important than REF for NRB in TKS roots. These findings would not only deepen our understanding of the TKS root proteome, but also provide new evidence on the roles of these NRB-related proteins in the mature TKS roots.

scholarly journals Fatty Acid Composition of Eucommia Rubber Particles, Content and Molecular Weight Distribution of Eu-rubber in Eucommia Ulmoides Leaves and Samaras

2020 ◽  
Author(s):  
Li-Jun Qin ◽  
Ting Yuan ◽  
ZhengWei Yang ◽  
DeGang Zhao
Keyword(s):  
Molecular Weight ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Molecular Weight Distribution ◽  
Acid Composition ◽  
Weight Distribution ◽  
Lipid Membrane ◽  
Eucommia Ulmoides ◽  
Rubber Biosynthesis ◽  
Rubber Particles

Abstract [Backgroud] Eucommia ulmoides rubber particles (EuRPs) wrapped by a monolayer protein-lipid membrane are speculated to an important site for the biosynthesis of E. ulmoides rubber (Eu-rubber), and the types and contents of these protein and lipid varied in different E. ulmoides organs which might indicate the difference of Eu-rubber biosynthesis in EuRPs from different organs and tissues. In this paper, the morphological structure and fatty acid composition of EuRPs in leaves and samaras collected in October, 2019 were compared, and molecular weight (Mw) and molecular weight distribution (MWD) were also analyed by GPC system. [Results] The results showed that small rubber particles (SRPs, 1~4 μm) and medium rubber particles (MRPs, 4~7 μm) from leaves and samaras respectively accounted for 39%, 53% and 38%, 34%, while the amount of the large rubber particles (LRPs, 7~11 μm) were 23% and 13%, which indicated there was no significance in leaf and samara SRPs while the amount of MRPs and LRPs in leaf both were more than that of in samara. Synchronously, the determination results of fatty acids (FAs) compositon showed three of the 35 kinds FAs were uniquely detected in leaf rubber particles (LeRPs), which included eleven carbonic (C11:0), thirteen carbonic (C13:0) and arachidic acids (C20:0), while γ-linolenic acid (C18:3n6) was exclusively found in samara rubber particles (SaRPs) with a lower content. Additioanlly, five characteristic distribution peaks (CDPs) of Eu-rubber from leaves were detected while there were only 2-3 CDPs in samaras; Further, the MWD analysis suggested that the MWD of Eu-rubber in E. ulmoides samaras and leaves were 40.61 and 6.76 respectively, which indicated the more Eu-rubber molecules with various Mw existed in samara compared with those in leaf. [Conclusions] Together, we concluded that average size of LeRPs was bigger than SaRPs in October, and 3 kinds of saturated fatty acid (SAF) were unique to the leaves while only one of unsaturated fatty acid (UFA) was sole to SaRPs. Simultaneously, the Eu-rubber in samara in October possessed more rubber molecules with various molecular weights compared with the contemporaneous leaves.

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