Dataset Information

Silica-based solid-phase extraction of cross-linked nucleic acid-bound proteins.

ABSTRACT: Proteins interact with nucleic acids to regulate cellular functions. The study of these regulatory interactions is often hampered by the limited efficiency of current protocols to isolate the relevant nucleic acid-protein complexes. In this report, we describe a rapid and simple procedure to highly enrich cross-linked nucleic acid-bound proteins, referred to as "2C" for "complex capture." This method is based on the observation that silica matrix-based columns used for nucleic acid purification also effectively retain UV cross-linked nucleic acid-protein complexes. As a proof of principle, 2C was used to isolate RNA-bound proteins from yeast and mammalian Huh7 cells. The 2C method makes RNA labelling redundant, and specific RNA-protein interactions can be observed and validated by Western blotting. RNA-protein complexes isolated by 2C can subsequently be immunoprecipitated, showing that 2C is in principle compatible with sensitive downstream applications. We suggest that 2C can dramatically simplify the study of nucleic acid-protein interactions and benefit researchers in the fields of DNA and RNA biology.

SUBMITTER: Asencio C

PROVIDER: S-EPMC6054301 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Silica-based solid-phase extraction of cross-linked nucleic acid-bound proteins.

Asencio Claudio C Chatterjee Aindrila A Hentze Matthias W MW

Life science alliance 20180618 3

Proteins interact with nucleic acids to regulate cellular functions. The study of these regulatory interactions is often hampered by the limited efficiency of current protocols to isolate the relevant nucleic acid-protein complexes. In this report, we describe a rapid and simple procedure to highly enrich cross-linked nucleic acid-bound proteins, referred to as "2C" for "complex capture." This method is based on the observation that silica matrix-based columns used for nucleic acid purification ...[more]

PMID: 30035255

Similar Datasets

Project description:Rare earth elements (REEs) and their compounds are essential for rapidly developing modern technologies. These materials are especially critical in the area of green/sustainable energy; however, only very high-purity fractions are appropriate for these applications. Yet, achieving efficient REE separation and purification in an economically and environmentally effective way remains a challenge. Moreover, current extraction technologies often generate large amounts of undesirable wastes. In that perspective, the development of selective, reusable, and extremely efficient sorbents is needed. Among numerous ligands used in the liquid-liquid extraction (LLE) process, the diglycolamide-based (DGA) ligands play a leading role. Although these ligands display notable extraction performance in the liquid phase, their extractive chemistry is not widely studied when such ligands are tethered to a solid support. A detailed understanding of the relationship between chemical structure and function (i.e., extraction selectivity) at the molecular level is still missing although it is a key factor for the development of advanced sorbents with tailored selectivity. Herein, a series of functionalized mesoporous silica (KIT-6) solid phases were investigated as sorbents for the selective extraction of REEs. To better understand the extraction behavior of these sorbents, different spectroscopic techniques (solid-state NMR, X-ray photoelectron spectroscopy, XPS, and Fourier transform infrared spectroscopy, FT-IR) were implemented. The obtained spectroscopic results provide useful insights into the chemical environment and reactivity of the chelating ligand anchored on the KIT-6 support. Furthermore, it can be suggested that depending on the extracted metal and/or structure of the ligand and its attachment to KIT-6, different functional groups (i.e., C═O, N-H, or silanols) act as the main adsorption centers and preferentially capture targeted elements, which in turn may be associated with the different selectivity of the synthesized sorbents. Thus, by determining how metals interact with different supports, we aim to better understand the solid-phase extraction process of hybrid (organo)silica sorbents and design better extraction materials.

Dataset Information

Silica-based solid-phase extraction of cross-linked nucleic acid-bound proteins.

Publications

Silica-based solid-phase extraction of cross-linked nucleic acid-bound proteins.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets