122nd General Meeting of the KCS

Type Poster Presentation
Area Life Chemistry
Room No. Grand Ballroom
Time 10월 19일 (금요일) 11:00~12:30
Code LIFE.P-446
Subject Supra-blot: Detection of Proteins Using Ultrastable Synthetic Binding Pairs
Authors Gihyun Sung, Jaehwan Sim1, Junghyun Kim2, Kyunglock Kim3, An Jaeyeon4, Sungwan Kim5, Hyun-Woo Rhee6, Kyeng Min Park7,*, Kimoon Kim8,*
AMS, Pohang University of Science and Technology, Korea
1IBIO, Pohang University of Science and Technology, Korea
2Institute for Basic Science, Korea
3Department of Pathology, Massachusetts General Hospital, United States
4Department of Chemistry, POSTECH, Korea
5Pohang University of Science and Technology, Korea
6Department of Chemistry, Ulsan National Institute of Science and Technology, Korea
7Center for Self-assembly and Complexity, Institute for Basic Science, Korea
8Department of Chemistry, Pohang University of Science and Technology, Korea
Abstract Modification of proteins with chemical tags has become a promising technique for proteomics by detecting the tags with their binding partners. Biotin has been broadly used as a chemical tag since it is specifically recognized by streptavidin with high binding affinity. A type of western blotting with horseradish peroxidase (HRP) conjugated streptavidin provides detection of the biotin tagged proteins for protein analysis. Although this streptavidin-biotin binding pair is very useful in protein detection, it suffers from various intrinsic problems such as non-desired enzymatic degradation of streptavidin, and unwanted interruption of endogenously existing biotin and biotinylated proteins, causing false positive signals and reduced sensitivity of protein detection. To avoid these problems, here we report a supramolecular chemistry-based protein detection method, termed it as “Supra-blot” using ultrastable synthetic binding pairs, adamantantylammonium (AdA) and cucurbit[7]uril (CB[7]). Specific proteins in bacteria and mammalian cells were labeled with AdA derivatives as chemical tags using various biological means. CB[7]-conjugated horseradish peroxidase (CB[7]-HRP) recognized the AdA-labeled proteins using ultrastable host-guest interactions. The labeled proteins were successfully detected with amplified chemiluminescence signals generated by CB[7]-HRP upon treating H2O2 and luminol. These promising outcomes showed a great potential of the use of Supra-blot for direct detection of AdA-labeled proteins of interest in chemical biology and proteomics.
E-mail junghyun8808@ibs.re.kr