Fishing Out Cancer Regulators With Molecular Baits

Using an anti-cancer drug as a bait, researchers have successfully fished out cancer regulating histone deacetylase proteins.

AsianScientist (Mar. 13, 2017) – Researchers at the Institute for Basic Science (IBS) have used pumpkin-shaped molecules to isolate cancer-prone proteins from complex samples. Their findings, published in Angewandte Chemie, could also be used to label proteins in live cells.

Histone deacetylases (HDACs) are an important family of proteins that regulate gene expression; an altered expression or mutations of genes that encode HDACs can induce tumors to develop. Therefore HDACs are among the most promising therapeutic targets for cancer treatment and they have inspired researchers to study and develop HDAC inhibitors.

Suberanilohydroxamic acid–ammonium-adamantane (SAHA) is an anti-cancer drug that slows down cancer progression by inhibiting HDACs, thereby preventing the replication of cancerous cells. In the present study, IBS researchers have used SAHA as a molecular bait to capture HDAC with the help of a high affinity bead they developed in 2011.

“The goal of this study was to purify histone deacetylase, a protein that plays a biologically important role in cells and is closely related to disease mechanisms such as cancer,” explained Dr. James Murray, the first author of the paper.

The bead is made from cucurbit[7]urils (CB[7]), a family of pumpkin-shaped macrocycles. These molecules have received a lot of attention because their high-affinity host–guest chemistry provides a unique opportunity to develop non-covalent materials for a range of applications.

“The CB[7]-based enrichment strategy takes advantage of small molecules with exceptionally high binding affinity. In contrast, recombinant methods use larger molecules with lower binding affinity,” Murray added.

Firstly, the researchers labeled a protein of interest with a high-affinity guest for their bead and extracted it from cell lysate using CB[7] beads. The CB[7] beads enriched guest-labeled HDACs from a cell lysate, the team also reported that the method may be useful for enriching proteins labeled from live cells.

“We used SAHA as the molecular bait that finds HDAC proteins,” said Murray. “Our molecule also has a functional group capable of forming a permanent bond with the captured protein when irradiated with ultraviolet light. As a result, it was possible to successfully purify the HDAC from a complex sample.”

The CB[7] bead method replaces traditional streptavidin-biotin purification methods which have an inherent high background reactivity. The researchers say that the CB[7] system is compatible with other commonly used protein handling methods and will be useful for deep proteome mining.


The article can be found at: Murray et al. (2017) Enrichment of Specifically Labeled Proteins by an Immobilized Host Molecule.

———

Source: Institute for Basic Science.
Disclaimer: This article does not necessarily reflect the views of AsianScientist or its staff.

Asian Scientist Magazine is an award-winning science and technology magazine that highlights R&D news stories from Asia to a global audience. The magazine is published by Singapore-headquartered Wildtype Media Group.

Related Stories from Asian Scientist