Wheat Proteome Profiling by Mass Spectrometry

Protein spots from ‘Aroona’ and 12 ARILs were excised from 2-D gels; reduced; alkylated; digested with trypsin, chymotrypsin, and thermolysin; and then analyzed as previously described [40 (link)]. The protein spots from 10 standard wheat cultivars were excised from 2-D gels and digested in-gel with chymotrypsin. The digested peptides were subjected to nanoAcquity UPLC coupled with MS (Waters Synapt G1 HDMS, MA, USA) to obtain their mass spectra. The data were analyzed using a database containing 63,245 Triticum protein sequences from NCBI (www.ncbi.nlm.nih.gov, 12 October 2015) and 151,173 Triticum protein sequences from UniProt (www.uniprot.org, 12 October 2015). UPLC–MS/MS analysis was conducted at the NICEM (National Instrumentation Center for Environmental Management, Seoul National University, Korea).