PXD008110

PXD008110 is an original dataset announced via ProteomeXchange.

Title	Efficiency of the purification protocols on complex matrices: biofilm, mouse liver and soil
Description	In gel-free experimental workflows, protein purification often starts with a precipitation stage using trichloroacetic acid (TCA). In presence of TCA, proteins precipitate in a stable molten globule state making the pellet difficultto solubilize in aqueous buffer for proteolytic digestion and MS analysis. In this context, the objective of this work was to study the suitability of a novel agent, ethanol/HCl, for the washing of TCA-precipitated proteins. This method optimized the recovery of proteins in aqueous buffer (50 to 96 %) while current organic solvents led to losses of material (up to 75 % of loss). Following a mechanistic study, the effect of ethanol/HCl on the conformation of TCA-precipitated proteins was investigated. It was shown that the reagent triggered the unfolding of TCA-stabilized molten globule into a reversible intermediate, characterized by a specific Raman signature, which favored protein subsequent resolubilization. Finally, the efficiency of ethanol/HCl for the washing of TCA-precipitated proteins extracted from biofilm, soil or mouse liver was demonstrated. Being versatile and simple, it could be of great interest to include an ethanol/HCl wash-step in mass spectrometry-based proteomics workflows to produce high-quality protein extracts.
HostingRepository	PRIDE
AnnounceDate	2017-11-30
AnnouncementXML	Submission_2018-07-27_08:44:14.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD008110
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Balkis EDDHIF
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Trichoderma cf. viride DIS 217I; NCBI TaxID: 493235; scientific name: Microchaete diplosiphon CCALA 811; NCBI TaxID: 1168853; scientific name: Porphyra purpurea; NCBI TaxID: 2787; scientific name: Synechocystis sp. CCALA 700; NCBI TaxID: 665890;
ModificationList	Carbamidomethyl
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-11-02 02:20:05	ID requested
1	2017-11-30 01:03:26	announced
2	2017-12-11 04:04:48	announced	Updated publication reference for PubMed record(s): 29191748.
⏵ 3	2018-07-27 08:44:15	announced	Updated publication reference for PubMed record(s): 29191748.

Eddhif B, Lange J, Guignard N, Batonneau Y, Clarhaut J, Papot S, Geffroy-Rodier C, Poinot P, Study of a novel agent for TCA precipitated proteins washing - comprehensive insights into the role of ethanol/HCl on molten globule state by multi-spectroscopic analyses. J Proteomics, 173():77-88(2018) [pubmed]

Eddhif B, Guignard N, Batonneau Y, Clarhaut J, Papot S, Geffroy-Rodier C, Poinot P, TCA precipitation and ethanol/HCl single-step purification evaluation: One-dimensional gel electrophoresis, bradford assays, spectrofluorometry and Raman spectroscopy data on HSA, Rnase, lysozyme - Mascots and Skyline data. Data Brief, 17():938-953(2018) [pubmed]

curator keyword: Technical

submitter keyword: biofilm, mouse liver, soil, LC-MS/MS

Pauline Poinot
contact affiliation	Institut de Chimie des Milieux et des Matériaux de Poitiers (IC2MP), Université de Poitiers, CNRS, Equipe Eaux,Biomarqueurs, Contaminants Organiques, Milieux, (E-BiCOM), 4 rue Michel Brunet, TSA 51106, F-86073 Poitiers, France.
contact email	pauline.poinot@univ-poitiers.fr
lab head
Balkis EDDHIF
contact affiliation	University of Poitiers
contact email	balkis.eddhif@univ-poitiers.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD008110
     1. Label: PRIDE project
     2. Name: Efficiency of the purification protocols on complex matrices: biofilm, mouse liver and soil