Chemical Sciences: A Manual for CSIR-UGC National Eligibility Test for Lectureship and JRF/Tandem mass tags

Tandem mass tags (TMT) are chemical labels used for mass spectrometry (MS) based quantitation 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 quantitation but may also be used in combination with these and other methods.

Versions

There are currently three varieties of TMT available:

• TMTzero - a non-isotopically substituted core structure
• TMTduplex - an isobaric pair of mass tags with a single isotopic substitution
• TMTsixplex - an isobaric set of six mass tags with five isotopic substitutions

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 quantitation 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.

Bioinformatics Integration

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 quantitate using TMT and other isobaric mass tags without the use of additional software.