Proteomic analysis of organ-specific post-translational lysine-acetylation and -methylation in mice by use of anti-acetyllysine and -methyllysine mouse monoclonal antibodies.
Post-translational lysine-acetylation and -methylation are two major PTMs of lysine residues in proteins. Recently, we established pan-reactive anti-acetyllysine mouse mAbs, which can bind to Nepsilon-acetylated lysine residues in various contexts of amino acid sequences. In the present study, we established pan-reactive anti-methyllysine mouse mAbs comparable to the anti-acetyllysine ones. By using these anti-acetyllysine and -methyllysine antibodies, we found that the pattern of lysine-acetylated and -methylated proteins in mouse organs showed extreme variation from organ to organ. We selected brain and skeletal muscle as model cases to be further analyzed by 2-DE followed by Western blotting. In brain, alpha-tubulin at its basal level was found to be extremely acetylated; and alpha-enolase was shown to be a newly recognized possibly acetylated protein. NF-L protein, Hsc70, alpha-tubulin fragments, beta-actin, and brain-type creatine kinase were identified as putative lysine-methylated proteins in mouse brain. In skeletal muscle, lysine-methylation of alpha-actin and both lysine-acetylation and -methylation of muscle-type creatine kinase were found as novel putative lysine-modified proteins. The approach presented here might be useful to find novel disease markers and/or drug target molecules that would not be noticed by use of the traditional proteomic approach only.
Iwabata
Hisako
H
R&D Division, Advanced Life Science Institute, 2-10-23 Maruyamadai, Wako, Saitama 351-0112, Japan.
Yoshida
Minoru
M
Komatsu
Yasuhiko
Y
D000107:Acetylation; D000199:Actins / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001923:Brain Chemistry; D052283:Creatine Kinase, MM Form / Q000032:analysis; D003429:Cross Reactions; D005260:Female; D006657:Histones / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D008239:Lysine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism*; D008745:Methylation; D051379:Mice; D009124:Muscle Proteins / Q000737:chemistry; D018482:Muscle, Skeletal / Q000737:chemistry; D009419:Nerve Tissue Proteins / Q000737:chemistry; D009928:Organ Specificity; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D040901:Proteomics; D014404:Tubulin / Q000378:metabolism
D000199:Actins; D000911:Antibodies, Monoclonal; D006657:Histones; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D014404:Tubulin; C005205:epsilon-N-methyllysine; C002315:N(alpha)-acetyllysine; C049907:2-methyllysine; C016949:N-epsilon-acetyllysine; D006735:Horseradish Peroxidase; D052283:Creatine Kinase, MM Form; D008239:Lysine