Residue-specific mass signatures for the efficient detection of protein modifications by mass spectrometry.
Currently available mass spectrometric (MS) techniques lack specificity in identifying protein modifications because molecular mass is the only parameter used to characterize these changes. Consequently, the suspected modified peptides are subjected to tandem MS/MS sequencing that may demand more time and sample. We report the use of stable isotope-enriched amino acids as residue-specific "mass signatures" for the rapid and sensitive detection of protein modifications directly from the peptide mass map (PMM) without enrichment of the modified peptides. These mass signatures are easily recognized through their characteristic spectral patterns and provide fingerprints for peptides containing the same content of specific amino acid residue(s) in a PMM. Without the need for tandem MS/MS sequencing, a peptide and its modified form(s) can readily be identified through their identical fingerprints, regardless of the nature of modifications. In this report, we demonstrate this strategy for the detection of methionine oxidation and protein phosphorylation. More interestingly, the phosphorylation of a histone protein, H2A.X, obtained from human skin fibroblast cells, was effectively identified in response to low-dose radiation. In general, this strategy of residue-specific mass tagging should be applicable to other posttranslational modifications.
Zhu
Haining
H
C-ACS, Chemistry Division, Los Alamos National Laboratory, New Mexico 87544, USA.
Hunter
Thomas C
TC
Pan
Songqin
S
Yau
Peter M
PM
Bradbury
E Morton
EM
Chen
Xian
X
D005347:Fibroblasts / Q000737:chemistry / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000737:chemistry
D006657:Histones; D011506:Proteins; D008715:Methionine