Correlation between phosphorylation ratios by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis and radioactivities by radioactive assay

To investigate the correlation between the counts per minute (CPM) by radioactivity assay and the phosphorylation ratio by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF MS) analysis, we prepared 136 peptide substrates. The correlation coefficient of phosphor...

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Bibliographic Details
Published in:Analytical biochemistry 2012-02, Vol.421 (2), p.773-775
Main Authors: Tsuchiya, Akira, Asai, Daisuke, Kang, Jeong-Hun, Mori, Takeshi, Niidome, Takuro, Katayama, Yoshiki
Format: Article
Language:English
Subjects:
amino acids
correlation
Correlation coefficient
desorption
ionization
MALDI–TOF MS
matrix-assisted laser desorption-ionization mass spectrometry
monitoring
peptides
Peptides - chemistry
Phosphorylation
Protein kinase
Radioactivity
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
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Summary:To investigate the correlation between the counts per minute (CPM) by radioactivity assay and the phosphorylation ratio by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF MS) analysis, we prepared 136 peptide substrates. The correlation coefficient of phosphorylation ratios to CPM was 0.77 for all samples. However, the more the numbers of positively charged amino acids increased, the more the correlation coefficient increased. Although positively charged amino acids can have an effect on the correlation results, MALDI–TOF MS analysis is a useful means for monitoring phosphorylated peptide and protein kinase activity instead of radioactivity assays.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2011.08.035