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-435
Subject Supramolecular Proteomic Mapping: Enrichment of Nuclear Proteins by Cucurbit[7]uril-immobilized Beads.
Authors Jaehwan Sim, Kyung Lock Kim1, Gihyun Sung2, Kyeng Min Park3,*, Kimoon Kim4,*
IBIO, Pohang University of Science and Technology, Korea
1Department of Pathology, Massachusetts General Hospital, United States
2AMS, Pohang University of Science and Technology, Korea
3Center for Self-assembly and Complexity, Institute for Basic Science, Korea
4Department of Chemistry, Pohang University of Science and Technology, Korea
Abstract Enrichment of proteins of interest is one of a key step in proteomic workflow. Currently, chemical labeling of a certain protein/protein family with biotin (Bt) and following enrichment with streptavidin (SA)-conjugated beads are widely used. However, it has inherent limitations such as biomolecular contamination from endogenously biotinylated proteins, damage of proteins in harsh eluting condition. Cucurbit[n]urils (CB[n]) are emerging macrocyclic molecules which can form host-guest binding pairs with high binding affinity comparable to the Bt-SA pair. We have reported a protein enrichment tool using cucurbit[7]uril (CB[7]) and ferrocenyl- (FcA) or adamantly-ammonium (AdA) as ultrastable host-guest binding pairs. Uses of the CB[7]-conjugated bead enabled us to enrich plasma membrane proteins and intracellular target proteins. To verify CB[7]-based host-guest binding pairs as a versatile proteomics tool, here, we report enrichment of nuclear proteins by combining the CB[7]-bead enrichment system with an enzymatic chemical tag labeling approach (APEX), a spatiotemporal protein labeling method. It enables us to isolate nuclear proteins with minimal biomolecular contamination and conserved spatiotemporal context. By achieving that, this supramolecular tool may become a powerful proteomics tool for isolation and enrichment of almost any protein located in cells, from the plasma membrane to the nucleus.
E-mail jhsim@postech.ac.kr