Identification of human, rat and chicken ribosomal proteins by a combination of two-dimensional polyacrylamide gel electrophoresis and mass spectrometry
To identify the exact spot position of human, rat and chicken ribosomal proteins (RP) separated by two-dimensional polyacrylamide gel electrophoresis (2-DE), a 2-DE system was designed to separate
RP with a pI > 8.6 according to their charge in the first dimension and to their molecular mass in the
second dimension. Individual proteins were excised from the gels and identified by mass spectrometry
after digestion by trypsin. In addition, a mixture of purified RP from these three species was also
analyzed by Tandem Mass Tag Spectrometry. By combining those two methods 74 RP from human,
76 from rat and 67 from chicken were identified according to the nomenclature initially defined for rat
liver RP and by using the Swiss-Prot/trEMBL databases. Whereas human and rat RP were well
described, most of RP from chicken were not characterized in databases, since 35 out of 67 chicken
RP identified in this study were not listed yet. We propose here the first comprehensive description of
chicken RP and their comparison to those from human and rat.
Foreskin (Cellntec), Rat Wistar (Cellntec)
Cultivation for cell fractional and ribosome purification, dimensional polyacrylamide gel electrophoresis and mass spectrometry of ribosomal proteins, their identification and relative quantification
