Gel-free based nano LC-MS/MS analysis of green rice leafhopper saliva
Saliva was collected from diet reared leafhoppers, pooled and concentrated. Samples were coming from Nephotettix cincticeps.
Sample Processing Protocol
Alkylated proteins were digested with trypsin and lysyl endopeptidase. Peptides were analyzed using nanospray LTQ XL Orbitrap MS. Protein identification from the MS/MS data was performed using using MASCOT software (Matrix Science) to correlate the data against N.cincticeps salivary gland RNA-seq data with 51,788 contigs (DDBJ Bio-Project Database ID PRJDB1562,) available at ftp://ftp.ddbj.nig.ac.jp/ddbj_database/dra/fastq/DRA002/DRA002228/DRZ003173/provisional/).
Data Processing Protocol
The acquired raw data files were processed using Proteome Discoverer (version 1.4, Thermo Fisher Scientific), and the parameters used in Mascot searches were as follows: carbamidomethylation of cysteine was set as a fixed modification and oxidation of methionine was set as a variable modification. Trypsin was specified as the proteolytic enzyme and one missed cleavage was allowed. Peptide mass tolerance was set at 5 ppm, fragment mass tolerance was set at 0.5 Da, and peptide charge was set at +2, +3, and +4. An automatic decoy database search was performed as part of the search.
Hattori M, Komatsu S, Noda H, Matsumoto Y. Proteome Analysis of Watery Saliva Secreted by Green Rice Leafhopper, Nephotettix cincticeps. PLoS One. 2015 Apr 24;10(4):e0123671. eCollection 2015 PubMed: 25909947