Abstract
A novel mass spectrometry-based assay system for determining protein kinase activity employing mass-tagged substrate peptide probes was used for the diagnosis of tumors. Two peptide probes (H-type and D-type) were synthesized containing the same substrate peptide sequence for protein kinase C (PKC). The molecular weights of the two probes differ because of the incorporation of deuterium into the acetyl groups of the D-type probe. The lysates of the normal and tumor tissue were prepared and reacted with the H- and D-type peptide probes, respectively. The PKC activities of the normal and tumor tissues can be compared simply and directly by calculating the phosphorylated ratio to each peptide probe, obtained from the peak intensity of the mass spectrum after mixing of the two reaction solutions. The phosphorylation ratio for the reaction of the H-type peptide probe with the tumor tissue lysate (B16 melanoma) was more than three times higher than that of the D type peptide probe with the normal skin tissue lysate. These results show that the novel assay system for detecting protein kinase activity using mass-tag technology can be a simple and useful means to profile protein kinase activity for cell or tissue lysate samples, and can be applied to the diagnosis of tumors.
| Original language | English |
|---|---|
| Pages (from-to) | 106-112 |
| Number of pages | 7 |
| Journal | Journal of the American Society for Mass Spectrometry |
| Volume | 18 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jan 1 2007 |
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All Science Journal Classification (ASJC) codes
- Structural Biology
- Spectroscopy
Cite this
Mass-Tag Technology Responding to Intracellular Signals as a Novel Assay System for the Diagnosis of Tumor. / Kang, Jeong Hun; Katayama, Yoshiki; Han, Aishan; Shigaki, Shuhei; Oishi, Jun; Kawamura, Kenji; Toita, Riki; Han, Xiao Ming; Mori, Takeshi; Niidome, Takuro.
In: Journal of the American Society for Mass Spectrometry, Vol. 18, No. 1, 01.01.2007, p. 106-112.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mass-Tag Technology Responding to Intracellular Signals as a Novel Assay System for the Diagnosis of Tumor
AU - Kang, Jeong Hun
AU - Katayama, Yoshiki
AU - Han, Aishan
AU - Shigaki, Shuhei
AU - Oishi, Jun
AU - Kawamura, Kenji
AU - Toita, Riki
AU - Han, Xiao Ming
AU - Mori, Takeshi
AU - Niidome, Takuro
PY - 2007/1/1
Y1 - 2007/1/1
N2 - A novel mass spectrometry-based assay system for determining protein kinase activity employing mass-tagged substrate peptide probes was used for the diagnosis of tumors. Two peptide probes (H-type and D-type) were synthesized containing the same substrate peptide sequence for protein kinase C (PKC). The molecular weights of the two probes differ because of the incorporation of deuterium into the acetyl groups of the D-type probe. The lysates of the normal and tumor tissue were prepared and reacted with the H- and D-type peptide probes, respectively. The PKC activities of the normal and tumor tissues can be compared simply and directly by calculating the phosphorylated ratio to each peptide probe, obtained from the peak intensity of the mass spectrum after mixing of the two reaction solutions. The phosphorylation ratio for the reaction of the H-type peptide probe with the tumor tissue lysate (B16 melanoma) was more than three times higher than that of the D type peptide probe with the normal skin tissue lysate. These results show that the novel assay system for detecting protein kinase activity using mass-tag technology can be a simple and useful means to profile protein kinase activity for cell or tissue lysate samples, and can be applied to the diagnosis of tumors.
AB - A novel mass spectrometry-based assay system for determining protein kinase activity employing mass-tagged substrate peptide probes was used for the diagnosis of tumors. Two peptide probes (H-type and D-type) were synthesized containing the same substrate peptide sequence for protein kinase C (PKC). The molecular weights of the two probes differ because of the incorporation of deuterium into the acetyl groups of the D-type probe. The lysates of the normal and tumor tissue were prepared and reacted with the H- and D-type peptide probes, respectively. The PKC activities of the normal and tumor tissues can be compared simply and directly by calculating the phosphorylated ratio to each peptide probe, obtained from the peak intensity of the mass spectrum after mixing of the two reaction solutions. The phosphorylation ratio for the reaction of the H-type peptide probe with the tumor tissue lysate (B16 melanoma) was more than three times higher than that of the D type peptide probe with the normal skin tissue lysate. These results show that the novel assay system for detecting protein kinase activity using mass-tag technology can be a simple and useful means to profile protein kinase activity for cell or tissue lysate samples, and can be applied to the diagnosis of tumors.
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UR - http://www.scopus.com/inward/citedby.url?scp=33845962371&partnerID=8YFLogxK
U2 - 10.1016/j.jasms.2006.09.004
DO - 10.1016/j.jasms.2006.09.004
M3 - Article
C2 - 17046276
AN - SCOPUS:33845962371
VL - 18
SP - 106
EP - 112
JO - Journal of the American Society for Mass Spectrometry
JF - Journal of the American Society for Mass Spectrometry
SN - 1044-0305
IS - 1
ER -