Indirect ultrasonication for protein quantification and peptide mass mapping through mass spectrometry-based techniques.
We report in this work a fast protocol for protein quantification and for peptide mass mapping that rely on (18)O isotopic labeling through the decoupling procedure. It is demonstrated that the purity and source of trypsin do not compromise the labeling degree and efficiency of the decoupled labeling reaction, and that the pH of the labeling reaction is a critical factor to obtain a significant (18)O double labeling. We also show that the same calibration curve can be used for MALDI protein quantification during several days maintaining a reasonable accuracy, thus simplifying the handling of the quantification process. In addition we demonstrate that (18)O isotopic labeling through the decoupling procedure can be successfully used to elaborate peptide mass maps. BSA was successfully quantified using the same calibration curve in different days and plasma from a freshwater fish, Cyprinus carpio, was used to elaborate the peptide mass maps.
Carreira
R J
RJ
REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516 Monte de Caparica, Portugal.
Lodeiro
C
C
Reboiro-Jato
M
M
Glez-Peña
D
D
Fdez-Riverola
F
F
Capelo
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D014465:Ultrasonics
D011506:Proteins