Methods of detecting protein arginine methyltransferase, and uses related thereto
AssigneeMDS Proteomics Inc.
InventorsCynthia J. Brame, Linda McBroom
Life of this patent
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Abstract
This invention relates to methods and reagents for detecting the presence of symmetrically and asymmetrically methylated arginine residues, either along or in polypeptides, using mass spectrometry. It also provides methods of determining structures of methylated arginine residues, methods of identifying substrates of PMRTs, and methods of conducting proteomic business using any of the suitable methods recited above.
Claims7
A method for identifying the structure of a dimethylarginine comprising:
- (i) obtaining, by mass spectroscopy, a neutral loss spectrum of a peptide containing a dimethylarginine, and
- (ii) ascertaining if the neutral loss spectrum shows one or both of neutral loss of monomethylamine (MMA), dimethylcarbodiimide (DMC), and/or neutral loss of dimethylarginine (DMA),
- wherein neutral loss of monomethylamine (MIMA) and of dimethylcarbodiimide (DMC) indicates the presence of a symmetrically dimethylated arginine residue, whereas neutral loss of dimethylamine (DMA) indicates the presence of an asymmetrically dimethylated arginine residue.
A method for identifying dimethylarginine residues in a peptide, comprising:
- (i) identifying the presence of dimethylarginine residues from mass spectra of said peptide obtained under conditions in which the spectra reveal mass modification by methylation, if any, of arginine residues in said peptide;
- (ii) ascertaining the nature of the methylation of a dimethylarginine residue identified in step (i) by determining if a neutral loss spectrum of said peptide shows one or both of neutral loss of monomethylamine (MMA), dimethylcarbodiimide (DMC), and/or neutral loss of dimethylamine (DMA), wherein neutral loss of monomethylamine (MMA) and of dimethylcarbodiimide (DMC) indicates the presence of a symmetrically dimethylated arginine residue, whereas neutral loss of dimethylamine (DMA) indicates the presence of a asymmetrically dimethylated arginine residue.
A method for identifying substrates of a protein arginine methyltransferase comprising:
- (i) obtaining mass spectra for one or more sample polypeptides which have been exposed to a protein arginine methyltransferase (PRMT) under conditions wherein methylation, if any, of arginine residues in said one or more sample polypeptides;
- (ii) identifying the presence of dimethylarginine residues in said one or more sample polypeptides by the presence of mass modified arginine in the mass spectra;
- and
- (iii) ascertaining the nature of the methylation of a dimethylarginine residue identified in step (i) by determining if a neutral loss spectrum of said one or more sample polytides shows one or both of neutral loss of monomethylamine (MMA), dimethylcarbodiimide (DMC), and/or neutral loss of dimethylamine (DMA), wherein neutral loss of monomethylamine (MMA) and of dimethylcarbodiimide (DMC) indicates the presence of a symmetrically dimethylated arginine residue whereas neutral loss of dimethylamine (DMA) indicates the presence of a asymmetrically dimethylated arginine residue.
Dependencies are inferred from the claim text.