TY - JOUR
T1 - Effect of the Addition of Diammonium Citrate to α-Cyano-4-Hydroxycinnamic Acid (CHCA) Matrix for the Detection of Phosphorylated Peptide in Phosphorylation Reactions Using Cell and Tissue Lysates
AU - Kang, Jeong Hun
AU - Toita, Riki
AU - Oishi, Jun
AU - Niidome, Takuro
AU - Katayama, Yoshiki
N1 - Funding Information:
This work was supported by CREST, Japan Science and Technology Corporation, a grant-in-aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture in Japan, and a grant-in-aid for Scientific Research from the Ministry of Health, Labor, and Welfare.
PY - 2007/11
Y1 - 2007/11
N2 - The ionization of phosphorylated peptides is usually suppressed by non-phosphorylated peptides when α-cyano-4-hydroxycinnamic acid (CHCA) is used as a matrix for matrix-assisted laser desorption/ionization-time-of-Flight (MALDI-TOF) mass spectrometry analysis. In the present study, we examined the effect of diammonium citrate addition to the CHCA matrix on the detection of phosphorylated peptides. Substrates for protein kinase C (PKC) and c-Src were synthesized and phosphorylated by reaction with cell and tissue lysate samples. The addition of diammonium citrate to the CHCA matrix increased the sensitivity for distinguishing phosphorylated peptides from background noise. However, the effect depended on substrate concentration.
AB - The ionization of phosphorylated peptides is usually suppressed by non-phosphorylated peptides when α-cyano-4-hydroxycinnamic acid (CHCA) is used as a matrix for matrix-assisted laser desorption/ionization-time-of-Flight (MALDI-TOF) mass spectrometry analysis. In the present study, we examined the effect of diammonium citrate addition to the CHCA matrix on the detection of phosphorylated peptides. Substrates for protein kinase C (PKC) and c-Src were synthesized and phosphorylated by reaction with cell and tissue lysate samples. The addition of diammonium citrate to the CHCA matrix increased the sensitivity for distinguishing phosphorylated peptides from background noise. However, the effect depended on substrate concentration.
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U2 - 10.1016/j.jasms.2007.08.002
DO - 10.1016/j.jasms.2007.08.002
M3 - Article
C2 - 17869536
AN - SCOPUS:35448994242
SN - 1044-0305
VL - 18
SP - 1925
EP - 1931
JO - Journal of the American Society for Mass Spectrometry
JF - Journal of the American Society for Mass Spectrometry
IS - 11
ER -