Generation of polyclonal antiserum for the detection of methylarginine proteins.

摘要: This report describes an approach for the study of biology methylarginine proteins based on generation immunological reagents capable recognizing status cellular proteins. Two forms immunizing peptide were prepared upon amino acid sequence motif found most prevalently among verified dimethylarginine-containing One form was constructed with 7 arginine residues alternating 8 glycine residues. None arginines used in synthesis methylated. The alternative synthesized identical repeating GRG sequence, but asymmetrical dimethylarginine at each residue. A methylarginine-specific antiserum generated using latter peptide. ELISA and western blotting arginine-rich peptides, or without asymmetric dimethylarginine, demonstrate methyl specificity antiserum. antibody co-localizes highly methylated native nucleolin protein conspicuously concentrated nucleolus. recognizes a bacterially expressed RBP16 only after incubation recombinant methyltransferase 1, cell extracts, respectively. Proteins isolated from cells different developmental states exhibit patterns reactivity observed by blots. Finally, reagent interacts specifically hnRNPA1 human fragile X mental retardation cultured PC12 cells. An detecting methylproteins will facilitate molecular analysis methylation wide variety research biomedical applications.