Tandem mass tag

Tandem mass tags (TMT or TMTs) are chemical labels used for mass spectrometry (MS)-based quantification and identification of biological macromolecules such as proteins, peptides and nucleic acids. TMT belongs to a family of reagents referred to as isobaric mass tags. They provide an alternative to gel- or antibody-based quantification but may also be used in combination with these and other methods.[1]

Contents

Versions

There are currently three varieties of TMT available:

The tags contain four regions, namely a mass reporter region (M), a cleavable linker region (F), a mass normalization region (N) and a protein reactive group (R). The chemical structures of all the tags are identical but each contains isotopes substituted at various positions, such that the mass reporter and mass normalization regions have different molecular masses in each tag. The combined M-F-N-R regions of the tags have the same total molecular weights and structure so that during chromatographic or electrophoretic separation and in single MS mode, molecules labelled with different tags are indistinguishable. Upon fragmentation in MS/MS mode, sequence information is obtained from fragmentation of the peptide back bone and quantification data are simultaneously obtained from fragmentation of the tags, giving rise to mass reporter ions.

Proteome Sciences Plc have the patents for TMT technology. These patents have broad coverage of all tandem mass tag technology.

Identification and quantification of TMT-labeled peptides

The structures of TMT tags are publicly available through the unimod database at unimod.org and hence, mass spectrometry software such as Mascot are able to account for the tag masses. Additionally, as of version 2.2, Mascot has the capability to quantify using TMT and other isobaric mass tags without the use of additional software.

See also

References

  1. ^ a b Thompson A, Schäfer J, Kuhn K et al. (2003). "Tandem mass tags: a novel quantification strategy for comparative analysis of complex protein mixtures by MS/MS". Anal. Chem. 75 (8): 1895–904. doi:10.1021/ac0262560. PMID 12713048. 
  2. ^ Dayon L, Hainard A, Licker V, Turck N, Kuhn K, Hochstrasser DF, Burkhard PR, Sanchez JC (2008). "Relative quantification of proteins in human cerebrospinal fluids by MS/MS using 6-plex isobaric tags". Anal. Chem. 80 (8): 2921–31. doi:10.1021/ac702422x. PMID 18312001. 
Methods
Isobaric labeling (Tandem mass tags (TMT) • Isobaric tags for relative and absolute quantitation (iTRAQ)) • Isotope-coded affinity tags (ICAT) • Label-free quantification (Spectral counting • Signal intensity) • Metal-coded tags (MeCATs) • N-terminal labeling • Stable isotope labeling by/with amino acids in cell culture (SILAC)