|
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 |
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