Plant Molecular Biology

, Volume 82, Issue 1, pp 155–168

In-depth proteome analysis of the rubber particle of Hevea brasiliensis (para rubber tree)

Article

DOI: 10.1007/s11103-013-0047-y

Cite this article as:
Dai, L., Kang, G., Li, Y. et al. Plant Mol Biol (2013) 82: 155. doi:10.1007/s11103-013-0047-y

Abstract

The rubber particle is a special organelle in which natural rubber is synthesised and stored in the laticifers of Hevea brasiliensis. To better understand the biological functions of rubber particles and to identify the candidate rubber biosynthesis-related proteins, a comprehensive proteome analysis was performed on H. brasiliensis rubber particles using shotgun tandem mass spectrometry profiling approaches—resulting in a thorough report on the rubber particle proteins. A total of 186 rubber particle proteins were identified, with a range in relative molecular mass of 3.9–194.2 kDa and in isoelectric point values of 4.0–11.2. The rubber particle proteins were analysed for gene ontology and could be categorised into eight major groups according to their functions: including rubber biosynthesis, stress- or defence-related responses, protein processing and folding, signal transduction and cellular transport. In addition to well-known rubber biosynthesis-related proteins such as rubber elongation factor (REF), small rubber particle protein (SRPP) and cis-prenyl transferase (CPT), many proteins were firstly identified to be on the rubber particles, including cyclophilin, phospholipase D, cytochrome P450, small GTP-binding protein, clathrin, eukaryotic translation initiation factor, annexin, ABC transporter, translationally controlled tumour protein, ubiquitin-conjugating enzymes, and several homologues of REF, SRPP and CPT. A procedure of multiple reaction monitoring was established for further protein validation. This comprehensive proteome data of rubber particles would facilitate investigation into molecular mechanisms of biogenesis, self-homeostasis and rubber biosynthesis of the rubber particle, and might serve as valuable biomarkers in molecular breeding studies of H. brasiliensis and other alternative rubber-producing species.

Keywords

Hevea brasiliensisRubber particleProteome1-DE gel electrophoresisMudPITMultiple reaction monitoringMass spectrometry

Abbreviations

REF

Rubber elongation factor

SRPP

Small rubber particle protein

CPT

cis-Prenyl transferase

IPP

Isopentenyl diphosphate

2-DE

Two-dimensional electrophoresis

SDS-PAGE

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

MS

Mass spectrometry

MS/MS

Tandem mass spectrometry

SCX

Strong cation exchange chromatography

MudPIT

Multidimensional protein identification technology

GeLC–MS

1D gel and liquid chromatography mass spectrometry

MRM

Multiple reaction monitoring

LC-MRM-MS

Liquid chromatography-MRM mass spectrometry

GO

Gene ontology

Mr

Relative molecular mass

Supplementary material

11103_2013_47_MOESM1_ESM.doc (78 kb)
Supplementary material 1 (DOC 77 kb)
11103_2013_47_MOESM2_ESM.xls (293 kb)
Supplementary material 2 (XLS 293 kb)
11103_2013_47_MOESM3_ESM.xls (286 kb)
Supplementary material 3 (XLS 286 kb)
11103_2013_47_MOESM4_ESM.fa (13.6 mb)
Supplementary material 4 (FA 13937 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.Key Laboratory of Biology and Genetic Resources of Rubber TreeMinistry of AgricultureDanzhouPeople’s Republic of China
  2. 2.State Key Laboratory Incubation Base, Rubber Research InstituteChinese Academy of Tropical Agricultural Sciences (CATAS)DanzhouPeople’s Republic of China