Protein complexes take DNA aptamer baits on a chip: A novel platform for detection of protein-protein interactions

ANYL 170

Lin-Chi Chen, chenlinchi@ntu.edu.tw1, Shin-Cheng Tzeng, newcity@ibms.sinica.edu.tw2, Wei-Chen Kao, kesley@gate.sinica.edu.tw2, and Konan Peck, konan@ibms.sinica.edu.tw2. (1) Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taipei, 10617, Taiwan, (2) Institute of Biomedical Sciences, Academia Sinica, Taipei, 115, Taiwan
By rivaling antibodies in highly specific and sensitive detection of target proteins, aptamers have been viewed as promising probes for protein chip applications. In this work, oligodeoxynucleotide aptamers (< 40 mer) were investigated as baits to capture protein complexes on a microarray surface. As proof of principle, we employed this method to detect possible interactions between thrombin and E. coli K12 total protein. From the chip assays with fluorescently labeled proteins, it was discovered that a thrombin-specific aptamer (15 mer) could associate a K12 protein only when thrombin was present. Furthermore, such association caused a competition effect that reduced protein binding signal to a K12-specific aptamer. After protein pull down assays, it was confirmed that E. coli Dps protein formed the complex with thrombin to take the bait aptamer on the chip. Accordingly, we consider an aptamer microarray a useful platform to simultaneously screen protein-protein interactions and bait ligands.
 

General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006