Supramolecular fishing for plasma membrane proteins using an ultrastable synthetic host–guest bindin
Author: ["Don-Wook Lee","Kyeng Min Park","Mainak Banerjee","Sang Hoon Ha","Taehoon Lee","Kyungwon Suh","Somak Paul","Hyuntae Jung","Jaeyoon Kim","Narayanan Selvapalam","Sung Ho Ryu","Kimoon Kim"]
Publication: Nature Chemistry
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Abstract
Membrane proteomics, the large-scale global analysis of membrane proteins, is often constrained by the efficiency of separating and extracting membrane proteins. Recent approaches involve conjugating membrane proteins with the small molecule biotin and using the receptor streptavidin to extract the labelled proteins. Despite the many advantages of this method, several shortcomings remain, including potential contamination by endogenously biotinylated molecules and interference by streptavidin during analytical stages. Here, we report a supramolecular fishing method for membrane proteins using the synthetic receptor–ligand pair cucurbit[7]uril–1-trimethylammoniomethylferrocene (CB[7]–AFc). CB[7]-conjugated beads selectively capture AFc-labelled proteins from heterogeneous protein mixtures, and AFc-labelling of cells results in the efficient capture of membrane proteins by these beads. The captured proteins can be recovered easily at room temperature by treatment with a strong competitor such as 1,1′-bis(trimethylammoniomethyl)ferrocene. This synthetic but biocompatible host–guest system may be a useful alternative to streptavidin–biotin for membrane proteomics as well as other biological and biotechnological applications. Characterization of plasma membrane proteins is important in understanding fundamental biological processes and developing new drugs, but their separation remains a challenge. Now, a synthetic receptor–ligand pair based on ferrocene derivatives and cucurbit[7]uril is shown to have exceptionally high binding affinity, and enables membrane proteins to be isolated efficiently without any contamination from naturally biotinylated molecules.
Cite this article
Lee, DW., Park, K., Banerjee, M. et al. Supramolecular fishing for plasma membrane proteins using an ultrastable synthetic host–guest binding pair. Nature Chem 3, 154–159 (2011). https://doi.org/10.1038/nchem.928
