Improved Protein Arrays for Quantitative Systems Analysis of the Dynamics of Signaling Pathway Interactions

YANG, CHIN-RANG¹ 

[1] NHLBI, NIH

关键词: protein array;    quantum dot;    quantitative analysis;    signaling pathway;    cross-talk;    image analysis;    systems biology;   

DOI  :  10.2172/1110247
RP-ID  :  DOE-UTSW-ER64335
PID  :  OSTI ID: 1110247

学科分类：放射科、核医学、医学影像

美国|英语

来源: SciTech Connect

【 摘 要 】

Astronauts and workers in nuclear plants who repeatedly exposed to low doses of ionizing radiation (IR, <10 cGy) are likely to incur specific changes in signal transduction and gene expression in various tissues of their body. Remarkable advances in high throughput genomics and proteomics technologies enable researchers to broaden their focus from examining single gene/protein kinetics to better understanding global gene/protein expression profiling and biological pathway analyses, namely Systems Biology. An ultimate goal of systems biology is to develop dynamic mathematical models of interacting biological systems capable of simulating living systems in a computer. This Glue Grant is to complement Dr. Boothman???s existing DOE grant (No. DE-FG02-06ER64186) entitled ???The IGF1/IGF-1R-MAPK-Secretory Clusterin (sCLU) Pathway: Mediator of a Low Dose IR-Inducible Bystander Effect??? to develop sensitive and quantitative proteomic technology that suitable for low dose radiobiology researches. An improved version of quantitative protein array platform utilizing linear Quantum dot signaling for systematically measuring protein levels and phosphorylation states for systems biology modeling is presented. The signals are amplified by a confocal laser Quantum dot scanner resulting in ~1000-fold more sensitivity than traditional Western blots and show the good linearity that is impossible for the signals of HRP-amplification. Therefore this improved protein array technology is suitable to detect weak responses of low dose radiation. Software is developed to facilitate the quantitative readout of signaling network activities. Kinetics of EGFRvIII mutant signaling was analyzed to quantify cross-talks between EGFR and other signaling pathways.