On-target titanium dioxide-based enrichment for characterization of phosphorylations in the Adenovirus pIIIa protein.
A recently developed titanium dioxide (TiO2) based on-target method for phosphopeptide enrichment and matrix assisted laser desorption-ionization mass spectrometry (MALDI MS) analysis was used to investigate phosphorylations in the Adenovirus type 2 structural protein pIIIa. Lysates of purified virus particles were separated on 1-D SDS-PAGE and the band for the pIIIa protein was excised for tryptic digestion into peptides that were enriched with the on-target method. The enrichment provided by the method clearly improved the detectability of phosphorylated peptides and the results show for the first time evidence for multi-phosphorylated peptides in pIIIa. Moreover, three novel phosphorylations were identified in the protein sequence, even though the precise positions could not be determined. These results illustrate the potential of the method for the characterization of novel phosphoproteomes in biological samples of medical relevance.
Department of Chemistry - BMC, Uppsala University, Uppsala, Sweden.
Anna I K
AI
Eriksson
Maik
M
Bartsch
Jonas
J
Bergquist
Katarina
K
Edwards
Sara Bergström
SB
Lind
Víctor
V
Agmo Hernández
D036022:Capsid Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D010748:Phosphopeptides / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D010766:Phosphorylation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014025:Titanium / Q000737:chemistry*; D014357:Trypsin
D036022:Capsid Proteins; D010446:Peptide Fragments; D010748:Phosphopeptides; C052227:protein IIIa, Human adenovirus type 2; C009495:titanium dioxide; D014025:Titanium; D014357:Trypsin
Capsid protein precursor pIIIa; MALDI-MS; Phosphopeptide enrichment; Separation; TiO(2)