Dataset Information

Mass spectrometry data confirming tetrameric ?-synuclein N-terminal acetylation.

ABSTRACT: Tetrameric ?-synuclein (?S) is an elusive multimer of the dynamic neuronal protein implicated in Parkinson?s disease. Through the data reported herein, we demonstrate that this high molecular weight multimer is N-acetylated. Coexpression of tetrameric ?S in Escherichia coli with the NatB acetylase derived from yeast enables access to N-terminally acetylated ?S (^NAc?S), the native form in humans. Following purification and characterization as previously described by us in "Isolation of Recombinant Tetrameric N-acetylated ?-synuclein" (Fernández and Lucas, 2018), the purified protein was excised from a native gel for confirmation of N-terminal acetylation. Through high-resolution mass spectrometry techniques, the identification of this helical tetramer as ^NAc?S has been clearly demonstrated.

SUBMITTER: Fernandez RD

PROVIDER: S-EPMC6157607 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Mass spectrometry data confirming tetrameric α-synuclein N-terminal acetylation.

Fernández Ricardo D RD Lucas Heather R HR

Data in brief 20180914

Tetrameric α-synuclein (αS) is an elusive multimer of the dynamic neuronal protein implicated in Parkinson׳s disease. Through the data reported herein, we demonstrate that this high molecular weight multimer is N-acetylated. Coexpression of tetrameric αS in <i>Escherichia coli</i> with the NatB acetylase derived from yeast enables access to N-terminally acetylated αS (<sup>NAc</sup>αS), the native form in humans. Following purification and characterization as previously described by us in "Isola ...[more]

PMID: 30263921

Similar Datasets

Project description:Histones are highly posttranslationally modified proteins that regulate gene expression by modulating chromatin structure and function. Acetylation and methylation are the most abundant histone modifications, with methylation occurring on lysine (mono-, di-, and trimethylation) and arginine (mono- and dimethylation) predominately on histones H3 and H4. In addition, arginine dimethylation can occur either symmetrically (SDMA) or asymmetrically (ADMA) conferring different biological functions. Despite the importance of histone methylation on gene regulation, characterization and quantitation of this modification have proven to be quite challenging. Great advances have been made in the analysis of histone modification using both bottom-up and top-down mass spectrometry (MS). However, MS-based analysis of histone posttranslational modifications (PTMs) is still problematic, due both to the basic nature of the histone N-terminal tails and to the combinatorial complexity of the histone PTMs. In this report, we describe a simplified MS-based platform for histone methylation analysis. The strategy uses chemical acetylation with d0-acetic anhydride to collapse all the differently acetylated histone forms into one form, greatly reducing the complexity of the peptide mixture and improving sensitivity for the detection of methylation via summation of all the differently acetylated forms. We have used this strategy for the robust identification and relative quantitation of H4R3 methylation, for which stoichiometry and symmetry status were determined, providing an antibody-independent evidence that H4R3 is a substrate for both Type I and Type II PRMTs. Additionally, this approach permitted the robust detection of H4K5 monomethylation, a very low stoichiometry methylation event (0.02% methylation). In an independent example, we developed an in vitro assay to profile H3K27 methylation and applied it to an EZH2 mutant xenograft model following small-molecule inhibition of the EZH2 methyltransferase. These specific examples highlight the utility of this simplified MS-based approach to quantify histone methylation profiles.

Dataset Information

Mass spectrometry data confirming tetrameric ?-synuclein N-terminal acetylation.

Publications

Mass spectrometry data confirming tetrameric α-synuclein N-terminal acetylation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets