Proteomics analysis of latex from Hevea brasiliensis (clone RRIM 600)

Abstract

The natural rubber latex extracted from the bark of Hevea brasiliensis plays various important roles in today's modern society. Following ultracentrifugation, the latex can be separated into 3 layers: C-serum, lutoids, and rubber particles. Previous studies have shown that a large number of proteins are present in these 3 layers. However, a complete proteome for this important plant is still unavailable. Protein sequences have been recently translated from the completed draft genome database of H. brasiliensis, leading to the creation of annotated protein databases of the following H. brasiliensis biosynthetic pathways: photosynthesis, latex allergens, rubberwood formation, latex biosynthesis, and disease resistance. This research was conducted to identify the proteins contained within the latex by way of de novo sequencing from mass spectral data obtained from the 3 layers of the latex. Peptides from these proteins were fragmented using collision-induced dissociation, higher-energy collisional dissociation, and electron-transfer dissociation activation methods. A large percentage of proteins from the biosynthetic pathways (63% to 100%) were successfully identified. In addition, a total of 1839 unique proteins were identified from the whole translated draft genome database (AnnHBM).

Identification and Characterization of Glycoproteins and Their Responsive Patterns upon Ethylene Stimulation in the Rubber Latex

“…It is known that rubber latex contains many macromolecular substances, such as proteins, sugars and alkaloids, and the NRB is a complex process [ 7 , 44 ]. Many proteins involved in the NRB process have been identified by proteomics-based technologies from different latex components [ 2 , 7 , 8 , 10 , 11 , 12 , 13 , 15 , 16 , 45 ]. These proteomics results revealed that rubber latex has a high protein content, and different kinds of PTMs have been determined in rubber latex to efficiently regulate NRB [ 7 , 8 ].…”

“…Phosphoglycerate mutase is known to exert a certain physiological impact in plant cells. It was identified as a metabolic protein from lettuce latex sap [ 47 ], rubber latex [ 13 ] and the washed solutions from rubber particles [ 48 ]. In this proteomics study, we found it was glycosylated (spot 3) in the 2-DE gels ( Figure 2 ) and increased by 2.3-fold after ethylene stimulation for three days ( Table S2 ).…”

“…Among the down-regulated ones, the genes encoding several enzymes known to be the key factors in rubber biosynthesis have been determined [ 5 ]. More recently published results also revealed that proteins might be the key regulators for NRB [ 10 , 11 , 12 , 13 , 14 ], and post-translational modifications (PTMs) of different protein isoforms may play crucial roles in NRB upon ethylene treatment [ 7 , 15 , 16 ], especially in small rubber particles [ 8 ]. There are more than three hundred kinds of reported PTMs in proteins [ 17 ].…”

Physiological and Proteomic Analyses of Molybdenum- and Ethylene-Responsive Mechanisms in Rubber Latex

“…Recently, many proteins had been identified by proteomics-based technologies from different Hevea latex components, such as total latex ( D’Amato et al, 2010 ; Li et al, 2011 ; Wang et al, 2015 ; Dai et al, 2016 ; Habib et al, 2017 ), rubber particles ( Rojruthai et al, 2010 ; Xiang et al, 2012 ; Dai et al, 2013 , 2016 ; Wang et al, 2016 ; Yamashita et al, 2016 ), C-serum ( Wagner et al, 2007 ; Wang et al, 2010 ; Havanapan et al, 2016 ), and lutoids ( Wang et al, 2010 , 2013 ; Habib et al, 2017 ). Proteins involved in mevalonate (MVA) pathway are crucial for NRB ( Wang et al, 2015 ).…”

Unrestrictive identification of post-translational modifications inHevea brasiliensislatex

“…All liquid chromatography-mass spectrometry (LC-MS) and tandem mass spectrometry (MS/MS) analyses were carried out according to our previous work (Habib et al 2016) by using Thermo LTQ/Orbitrap Velos coupled with Easy-nLC II system. Briefly, the tryptic-digested peptides chromatographic separation was performed using Easy-Column C18-A2 (100 0.75 mm i.d., 3 m) coupled with pre-column (Easy-Column, 20 0.1 mm i.d., 5 m) at flow rate of 0.3 l/min and injection volume of sample was 10 µl.…”