A system and method for automatic protein sequencing by mass...

Details A system and method for automatic protein sequencing by mass... A system and method for automatic protein sequencing by mass...

G - Physics

01

N

G01N 33/68 (2006.01) H01J 49/00 (2006.01) H01J 49/04 (2006.01)

Patent

CA 2468689

A method of deducing the sequence of a protein by analysing tandem mass spectrometry data. The protein is subjected to partial isotopic labelling by enzymatic digestion in a water mixture comprising a non-natural abundance of H21sO. Differential scanning mass spectrometry is applied to the peptide fragments obtained from the digestion. Peaks in the spectra are analyzed to ascertain whether they arise from isotopically labeled fragments or not. A filtered spectrum is calculated that just comprises peaks from the y-ions. The sequence of the peptide is deduced by computing the difference in the mass between adjacent y-ion peaks.

L'invention concerne un procédé permettant la déduction de la séquence d'une protéine à partir de l'analyse des données d'une spectrométrie de masse en tandem. Ce procédé consiste à appliquer un marquage isotopique partiel est appliqué à la protéine par digestion enzymatique dans un mélange aqueux présentant une teneur non naturelle en H¿2??18¿O. Les fragments peptidiques résultant de cette digestion sont soumis à une spectrométrie de masse à balayage différentiel. Une analyse des pics du spectre est ensuite effectuée afin de déterminer s'ils proviennent des fragments marqués par un isotope ou non. Un spectre filtré qui ne comprend que les pics provenant des ions y est alors calculé. La séquence du peptide est déduite à partir du calcul de la différence de masse entre les pics des ions y adjacents.

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