Abstract
An intracellular signal transduction system involved in many kinds of proteins regulates various cellular events, such as cellular proliferation, differentiation and cell death, responding to the outer environment. Among those proteins, protein kinases are one of the most important class of proteins. It has been reported that abnormal activities of protein kinases are related to various diseases. Therefore, an analysis of protein kinase activity would be useful for diagnosis and drug screening. Herein, we describe the detection of on-chip phosphorylation using a peptide chip by fluorescence imaging. In this work, we selected a cAMP-dependent protein kinase (PKA) as a model, and quantitatively evaluated the substrate specificity for PKA using a peptide chip. Moreover, we attempted to detect intracellular kinase activity. Since src correlates with breast cancer, we measured the src activity in MCF-7 human breast cancer cell lysate. Consequently, with our peptide chip we could detect the src activity in MCF-7 cell lysate. Therefore, this method could be applied to diagnosis, drug discovery and screening.
| Original language | English |
|---|---|
| Pages (from-to) | 969-973 |
| Number of pages | 5 |
| Journal | bunseki kagaku |
| Volume | 55 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - Dec 1 2006 |
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All Science Journal Classification (ASJC) codes
- Analytical Chemistry
Cite this
Phosphorylation signal detection on a peptide chip. / Shigaki, Shuhei; Han, Xiaoming; Yamaji, Takayuki; Yamanouchi, Go; Mori, Takeshi; Nidome, Takuro; Katayama, Yoshiki.
In: bunseki kagaku, Vol. 55, No. 12, 01.12.2006, p. 969-973.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Phosphorylation signal detection on a peptide chip
AU - Shigaki, Shuhei
AU - Han, Xiaoming
AU - Yamaji, Takayuki
AU - Yamanouchi, Go
AU - Mori, Takeshi
AU - Nidome, Takuro
AU - Katayama, Yoshiki
PY - 2006/12/1
Y1 - 2006/12/1
N2 - An intracellular signal transduction system involved in many kinds of proteins regulates various cellular events, such as cellular proliferation, differentiation and cell death, responding to the outer environment. Among those proteins, protein kinases are one of the most important class of proteins. It has been reported that abnormal activities of protein kinases are related to various diseases. Therefore, an analysis of protein kinase activity would be useful for diagnosis and drug screening. Herein, we describe the detection of on-chip phosphorylation using a peptide chip by fluorescence imaging. In this work, we selected a cAMP-dependent protein kinase (PKA) as a model, and quantitatively evaluated the substrate specificity for PKA using a peptide chip. Moreover, we attempted to detect intracellular kinase activity. Since src correlates with breast cancer, we measured the src activity in MCF-7 human breast cancer cell lysate. Consequently, with our peptide chip we could detect the src activity in MCF-7 cell lysate. Therefore, this method could be applied to diagnosis, drug discovery and screening.
AB - An intracellular signal transduction system involved in many kinds of proteins regulates various cellular events, such as cellular proliferation, differentiation and cell death, responding to the outer environment. Among those proteins, protein kinases are one of the most important class of proteins. It has been reported that abnormal activities of protein kinases are related to various diseases. Therefore, an analysis of protein kinase activity would be useful for diagnosis and drug screening. Herein, we describe the detection of on-chip phosphorylation using a peptide chip by fluorescence imaging. In this work, we selected a cAMP-dependent protein kinase (PKA) as a model, and quantitatively evaluated the substrate specificity for PKA using a peptide chip. Moreover, we attempted to detect intracellular kinase activity. Since src correlates with breast cancer, we measured the src activity in MCF-7 human breast cancer cell lysate. Consequently, with our peptide chip we could detect the src activity in MCF-7 cell lysate. Therefore, this method could be applied to diagnosis, drug discovery and screening.
UR - http://www.scopus.com/inward/record.url?scp=33845786926&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33845786926&partnerID=8YFLogxK
U2 - 10.2116/bunsekikagaku.55.969
DO - 10.2116/bunsekikagaku.55.969
M3 - Article
VL - 55
SP - 969
EP - 973
JO - Bunseki Kagaku
JF - Bunseki Kagaku
SN - 0525-1931
IS - 12
ER -