The biosynthesis of rubber: Incorporation of isopentenyl pyrophosphate into purified rubber particles by a soluble latex-serum enzyme

McMullen, A.I.; McSweeney, G. P.

doi:10.1042/bj1010042

Cited by 45 publications

(10 citation statements)

References 7 publications

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“…brusiliensis (Archer et al, 1963 ;McMullen and McSweeney, 1966). However, in common with the purified prenyl transferase , these other extracts could not synthesize rubber in solution; all had an absolute requirement for rubber particles, E elusticu washed rubber particles often being the particles of choice.…”

Section: Discussionmentioning

confidence: 99%

“…Rubber biosynthesis can occur in the absence of soluble proteins if these are substituted for by a number of different allylic diphosphate initiator molecules (Archer and Audley, 1987;Audley and Archer, 1988;Berndt, 1963;Cornish and Backhaus, 1990;Madhavan et al, 1989). Thus, it is probable that all soluble extracts reported to have rubber transferase activity (Archer et al, 1963;McMullen and McSweeney, 1966) were actually part of the rubber molecule initiation system, synthesizing essential allylic diphosphates but not cis-1,4-p0lyiso-prene.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

The separate roles of plant cis and trans prenyl transferases in cis‐1,4‐polyisoprene biosynthesis

Cornish¹

1993

European Journal of Biochemistry

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In plants, the elongation of cis-1,4-polyisoprene (natural rubber, M, > lo6) requires a small transallylic diphosphate ( S C2J initiator. The trans-allylic diphosphates are hydrophilic cytosolic compounds, whereas cis-l,4-polyisoprene is hydrophobic and compartmentalised in subcellular rubber particles.In this paper, it is demonstrated that soluble trans-prenyl transferase from latex of Hevea brusiliensis functions solely as farnesyl diphosphate synthase, and plays no direct role in cis-1,4-polyisoprene elongation. The cis-l,4-prenyl transferase is firmly associated with the H. bvasiliensis rubber particle, as is also the case in other rubber-producing species

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The separate roles of plant cis and trans prenyl transferases in cis‐1,4‐polyisoprene biosynthesis

Cornish¹

1993

European Journal of Biochemistry

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“…The amount of [1][2][3][4][5][6][7][8][9][10][11][12][13][14] C]IPP incorporation in the toluene/hexane extracts of 2500 dpm that was observed in the control reaction with WBP is attributable to the endogenous rubber transferase activity in the WBP. When the homogenates of BL21(DE3)/pETHRT1 were coincubated with WBP, the amount of IPP incorporation into the toluene/hexane extracts was similar to that of the control experiment with WBP alone.…”

Section: Enzymatic Activity Of Hrt2 Proteinsmentioning

confidence: 99%

Molecular cloning, expression and characterization of cDNA encoding cis‐prenyltransferases from Hevea brasiliensis

Asawatreratanakul

Zhang

Wititsuwannakul

et al. 2003

European Journal of Biochemistry

158

119

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Natural rubber from Hevea brasiliensis is a high molecular mass polymer of isoprene units with cis-configuration. The enzyme responsible for the cis-1,4-polymerization of isoprene units has been identified as a particle-bound rubber transferase, but no gene encoding this enzyme has been cloned from rubber-producing plants. By using sequence information from the conserved regions of cis-prenyl chain elongating enzymes that were cloned recently, we have isolated and characterized cDNAs from H. brasiliensis for a functional factor participating in natural rubber biosynthesis. Sequence analysis revealed that all of the five highly conserved regions among cis-prenyl chain elongating enzymes were found in the protein sequences of the Hevea cis-prenyltransferase. Northern blot analysis indicated that the transcript(s) of the Hevea cis-prenyltransferase were expressed predominantly in the latex as compared with other Hevea tissues examined. In vitro rubber transferase assays using the recombinant gene product overexpressed in Escherichia coli revealed that the enzyme catalyzed the formation of long chain polyprenyl products with approximate sizes of 2 · 10 3 )1 · 10 4 Da. Moreover, in the presence of washed bottom fraction particles from latex, the rubber transferase activity producing rubber product of high molecular size was increased. These results suggest that the Hevea cis-prenyltransferase might require certain activation factors in the washed bottom fraction particles for the production of high molecular mass rubber.

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“…Arg., clone RY 7‐33‐97) as previously described . After centrifugation at 40 000 × g for 60 min at 4°C, the upper fraction containing rubber particles was purified using a WS containing 10 mM EDTA Na 2 , 0.5 mM DTT, 250 mM Sucrose, and 20 mM Tris‐HCl, pH 7.0 as previously described . The used WS was collected for protein extraction.…”

Section: Methodsmentioning

confidence: 99%

A protein extraction method for low protein concentration solutions compatible with the proteomic analysis of rubber particles

et al. 2016

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The extraction of high-purity proteins from the washing solution (WS) of rubber particles (also termed latex-producing organelles) from laticifer cells in rubber tree for proteomic analysis is challenging due to the low concentration of proteins in the WS. Recent studies have revealed that proteins in the WS might play crucial roles in natural rubber biosynthesis. To further examine the involvement of these proteins in natural rubber biosynthesis, we designed an efficiency method to extract high-purity WS proteins. We improved our current borax and phenol-based method by adding reextraction steps with phenol (REP) to improve the yield from low protein concentration samples. With this new method, we extracted WS proteins that were suitable for proteomics. Indeed, compared to the original borax and phenol-based method, the REP method improved both the quality and quantity of isolated proteins. By repeatedly extracting from low protein concentration solutions using the same small amount of phenol, the REP method yielded enough protein of sufficiently high-quality from starting samples containing less than 0.02 mg of proteins per milliliter. This method was successfully applied to extract the rubber particle proteins from the WS of natural rubber latex samples. The REP-extracted WS proteins were resolved by 2DE, and 28 proteins were positively identified by MS. This method has the potential to become widely used for the extraction of proteins from low protein concentration solutions for proteomic analysis.

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The biosynthesis of rubber: Incorporation of isopentenyl pyrophosphate into purified rubber particles by a soluble latex-serum enzyme

Cited by 45 publications

References 7 publications

The separate roles of plant cis and trans prenyl transferases in cis‐1,4‐polyisoprene biosynthesis

The separate roles of plant cis and trans prenyl transferases in cis‐1,4‐polyisoprene biosynthesis

Molecular cloning, expression and characterization of cDNA encoding cis‐prenyltransferases from Hevea brasiliensis

A protein extraction method for low protein concentration solutions compatible with the proteomic analysis of rubber particles

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