Proteomic Identification of Hevea Proteins

After the target protein spots were cut out, the in-gel digestion was performed as described [8 (link)]. After enzymatic digestion, these peptides were collected and identified by the AB 5800 MALDI-TOF/TOF mass spectrometry (MS) instrument (AB SCIEX, Foster City, CA, USA). The ProteinPilot Software (Version 4.5) and a Mascot Algorithm (version 2.3) were used to search against the Hevea genome scaffolds (BioProject ID: PRJNA80191, www.ncbi.nlm.nih.gov/nuccore/448814761) and the draft genome (GenBank: AJJZ01000000) with 46,718 sequences and 17,435,757 residues [63 (link)]. The parameters were set as: precursor tolerance 300 ppm, 0.3 Da tolerance as MS/MS fragment, trypsin as the enzyme, carbamidomethylation as the fixed modification (C) and oxidation (M) as the variable modification. Detailed information on all identified protein spot mass searches can be found in Supplemental Figure S1 and Table S1.