Updated project metadata. Post-translational modification of proteins through methylation plays important regulatory role in biological processes. Lysine methylation on histone proteins is known to play important role in chromatin structure and function. However, non-histone protein substrates of this modification remain largely unknown. Herein, we use high resolution mass spectrometry to global screening methylated substrates and lysine- methylation sites in tomato (Solanum Lycopersicum). A total of 241 sites of lysine methylation (mono-, di-, tri-methylation) in 176 proteins with diverse biological functions and subcellular localized were identified in mix tomato with different maturity. Two putative methylation motifs were detected. KEGG pathway category enrichment analysis indicated that methylated proteins are implicated in the regulation of diverse metabolic processes, including arbon fixation in photosynthetic organisms, pentose phosphate pathway, fructose and mannose metabolism, and cysteine and methionine metabolism. Three representative proteins were selected to analyze the effect of methylated modification on protein function. In addition, quantitative RT-PCR further validated the gene expression level of some key methylated proteins during fruit ripening, which are involved in oxidation reduction process, stimulus and stress, energy metabolism, signaling transduction, fruit ripening and senescence. These data represent the first report of methylation proteomic and supply abundant resources for exploring the functions of lysine methylation in tomato and other plants.