TY - JOUR
T1 - Dual fluorescent molecular substrates selectively report the activation, sustainability and reversibility of cellular PKB/Akt activity
AU - Shen, Duanwen
AU - Bai, Mingfeng
AU - Tang, Rui
AU - Xu, Baogang
AU - Ju, Xiaoming
AU - Pestell, Richard G.
AU - Achilefu, Samuel
N1 - Funding Information:
We thank Mikhail Berezin and Sharon Bloch (Radiology) for useful discussions, as well as Fongfu Hsu and Alan N. Bohrer (Internal Medicine) for technical assistance with MS/MS analysis. This work was supported in part by grants from the National Institutes of Health (NIBIB R01 EB008111 and R01 EB008458; and NCI R01 CA171651) and the National Science Foundation (CCF 0963742). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
PY - 2013
Y1 - 2013
N2 - Using a newly developed near-infrared (NIR) dye that fluoresces at two different wavelengths (dichromic fluorescence, DCF), we discovered a new fluorescent substrate for Akt, also known as protein kinase B, and a method to quantitatively report this enzyme's activity in real time. Upon insulin activation of cellular Akt, the enzyme multi-phosphorylated a single serine residue of a diserine DCF substrate in a time-dependent manner, culminating in monophospho-to triphospho-serine products. The NIR DCF probe was highly selective for the Akt1 isoform, which was demonstrated using Akt1 knockout cells derived from MMTV-ErbB2 transgenic mice. The DCF mechanism provides unparalleled potential to assess the stimulation, sustainability, and reversibility of Akt activation longitudinally. Importantly, NIR fluorescence provides a pathway to translate findings from cells to living organisms, a condition that could eventually facilitate the use of these probes in humans.
AB - Using a newly developed near-infrared (NIR) dye that fluoresces at two different wavelengths (dichromic fluorescence, DCF), we discovered a new fluorescent substrate for Akt, also known as protein kinase B, and a method to quantitatively report this enzyme's activity in real time. Upon insulin activation of cellular Akt, the enzyme multi-phosphorylated a single serine residue of a diserine DCF substrate in a time-dependent manner, culminating in monophospho-to triphospho-serine products. The NIR DCF probe was highly selective for the Akt1 isoform, which was demonstrated using Akt1 knockout cells derived from MMTV-ErbB2 transgenic mice. The DCF mechanism provides unparalleled potential to assess the stimulation, sustainability, and reversibility of Akt activation longitudinally. Importantly, NIR fluorescence provides a pathway to translate findings from cells to living organisms, a condition that could eventually facilitate the use of these probes in humans.
UR - http://www.scopus.com/inward/record.url?scp=84877744045&partnerID=8YFLogxK
U2 - 10.1038/srep01697
DO - 10.1038/srep01697
M3 - Article
C2 - 23603888
AN - SCOPUS:84877744045
SN - 2045-2322
VL - 3
JO - Scientific reports
JF - Scientific reports
M1 - 1697
ER -