Top-down proteomics with a quadrupole time-of-flight mass spectrometer and collision-induced dissociation.
With slight modifications of the instrumental parameters, we demonstrate that satisfactory top-down data can be obtained with collision-induced dissociation (CID) tandem mass spectrometry on a quadrupole time-of-flight (qTOF) instrument not originally designed for this purpose. Protein identification is achieved with both N- and C-terminal sequence tags and BLAST database searches. The accurate mass measurement of multiply charged fragment ions (mostly y and b-type) supplements the limited set of cleavage sites and provides a high degree of sequence coverage (90-100%). Post-translational modification issues can be addressed too. This approach might help those mass spectrometry (MS) core facilities that are not able to afford very high-resolution instruments, thus expanding the benefits of top-down protein analysis over the worldwide MS community.
Armirotti
Andrea
A
Centro Biotecnologie Avanzate, Largo Rosanna Benzi 10, 16132 Genova, Italy. andrea.armirotti@unige.it
Benatti
Umberto
U
Damonte
Gianluca
G
D000595:Amino Acid Sequence; D000818:Animals; D024402:Carbonic Anhydrase II / Q000032:analysis / Q000737:chemistry*; D002417:Cattle; D020869:Gene Expression Profiling / Q000379:methods*; D006736:Horses; D008969:Molecular Sequence Data; D009211:Myoglobin / Q000032:analysis / Q000737:chemistry*; D020543:Proteome / Q000032:analysis / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D009211:Myoglobin; D020543:Proteome; D024402:Carbonic Anhydrase II