2-D Fluorescence Difference Gel Electrophoresis (DIGE) in Neuroproteomics

Díez, Roberto A.; Herbstreith, Michael H.; Osorio, Cristina; Álzate, Óscar

doi:10.1201/9781420076264.ch4

Cited by 13 publications

(12 citation statements)

References 75 publications

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“…For post-translational modification analysis, FLAG-tagged GPSM3 immunocomplex beads were submitted to the Systems Proteomics Center of University of North Carolina School (UNC)-Chapel Hill (directed by Dr. Oscar Alzate) and analyzed for post-translational modification using two-dimensional-differential in gel electrophoresis (23). Phosphorylation detected within GPSM3 was further analyzed using tandem mass spectrometry.…”

Section: Methodsmentioning

confidence: 99%

Regulation of the Subcellular Localization of the G-protein Subunit Regulator GPSM3 through Direct Association with 14-3-3 Protein

Giguère

Laroche

Oestreich

et al. 2012

Journal of Biological Chemistry

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Background: GPSM3, a guanine nucleotide dissociation inhibitor acting on G␣ i subunit family members, is both cytoplasmand nucleus-localized. Results: GPSM3 interacts directly with 14-3-3 through a phosphorylation-dependent mechanism. Conclusion: Interaction with 14-3-3 stabilizes a cytoplasmic pool of GPSM3. Significance: Our findings represent the first demonstration of post-translational modulation of GPSM3, a hematopoieticrestricted G-protein subunit regulator.

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Section: Methodsmentioning

confidence: 99%

Regulation of the Subcellular Localization of the G-protein Subunit Regulator GPSM3 through Direct Association with 14-3-3 Protein

Giguère

Laroche

Oestreich

et al. 2012

Journal of Biological Chemistry

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“…The first reports of untargeted proteomic large screenings in schizophrenia begun by using two-dimensional electrophoresis (2-DE) [30] where the first dimension is an isoelectric focusing step (IEF) followed by the second dimension which is a standard SDS-PAGE. In this approach the quantitative information is retrieved from the analysis of the intensity of the staining of a given spot in the gel, which is then picked, and the proteins are identified by MS. Later on, an improvement of this technology was introduced -two-dimensional difference gel electrophoresis (2D-DIGE) [31], where the separation steps remain the same but proteins are directly labelled with fluorescent dyes (CyDies), enabling the analysis of multiple samples in the same gel, this way improving the reproducibility of the quantitative results [32].…”

Section: Proteomics Contribution For Schizophrenia's Biomarker Researchmentioning

confidence: 99%

“…Although 2-DE techniques have been extremely important in transforming protein analysis into an "omics" approach, these methods have some drawbacks as the lengthy sample preparation and difficulty in detecting some subtypes of proteins, like membrane proteins, or due to dynamic range limitations [32]. Throughout the years researchers have turned to other methodologies that can measure quantitative levels of the entire set of proteins, and that can be divided into two main groups: (i) labelling techniques, which require the chemical, metabolic or enzymatic stable isotopic labelling of the samples prior to MS analysis; (ii) label-free techniques, which are gaining increasing interest due to improvements in accuracy and sensitivity of MS instruments and data processing algorithms [33].…”

Section: Proteomics Contribution For Schizophrenia's Biomarker Researchmentioning

confidence: 99%

Circulating biomarkers in schizophrenia: a proteomics perspective

Santa¹,

Coelho²,

Madeira³

et al. 2017

IJCNMH

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Schizophrenia (SCZ) is one of the most severe and devastating major mental disorders, with an onset typically in late adolescence or early adulthood affecting about 0.5-1.2% of the population worldwide. It is one of the most debilitating medical conditions, with striking socio-economic burden to society due to lost employment and social support that largely surpass direct costs of treatment. SCZ is listed by the World Health Organization (WHO) among the top 20 leading causes of disability worldwide. Its diagnosis is syndromic, based in clinical interviewing and anamnesis, as there is to date no biochemical test to aid in this diagnosis. On the other hand, SCZ treatment is mostly based in antipsychotic drugs which are in several aspects somehow ineffective, and some of these medications have low tolerability with severe side effects. For all these reasons, the current search for new panels of biomarkers is of the utmost importance to aid in the correct diagnosis and stratification of the patients, prognosis, and prediction of treatment effectiveness.In this review, a total of 25 publications on peripheral SCZ biomarkers are presented from proteomics studies performed in body fluids of patients searching for protein markers, and using mass spectrometry. To date such proteomics studies have already been achieved in cerebrospinal fluid (CSF), serum, plasma, peripheral blood cells (namely, mononuclear cells, T-cells and red blood cells), saliva, and sweat, being of paramount importance in the schizophrenia research field, but still lacking validation and clinical translation.In summary, a general overview of the results from these 25 studies, as well as the challenges and future perspectives of the field, are presented and discussed.

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“…Ten potential multiple sclerosis biomarkers were selected for validation by immunoassay (Ottervald et al, 2010). These methodologies, sample preparation techniques, and applications of 2D-DIGE in neuroproteomics were reviewed by Diez et al(Diez et al, 2010). Although 2D gel provides excellent resolving power and capability to visualize abundance changes, there are some limitations to the method.…”

Section: Fractionation and Separationmentioning

confidence: 99%

Mass spectrometry-based proteomics and peptidomics for systems biology and biomarker discovery

Cunningham

2012

Front. Biol.

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The scientific community has shown great interest in the field of mass spectrometry-based proteomics and peptidomics for its applications in biology. Proteomics technologies have evolved to produce large datasets of proteins or peptides involved in various biological and disease progression processes producing testable hypothesis for complex biological questions. This review provides an introduction and insight to relevant topics in proteomics and peptidomics including biological material selection, sample preparation, separation techniques, peptide fragmentation, post-translation modifications, quantification, bioinformatics, and biomarker discovery and validation. In addition, current literature and remaining challenges and emerging technologies for proteomics and peptidomics are presented.

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2-D Fluorescence Difference Gel Electrophoresis (DIGE) in Neuroproteomics

Cited by 13 publications

References 75 publications

Regulation of the Subcellular Localization of the G-protein Subunit Regulator GPSM3 through Direct Association with 14-3-3 Protein

Regulation of the Subcellular Localization of the G-protein Subunit Regulator GPSM3 through Direct Association with 14-3-3 Protein

Circulating biomarkers in schizophrenia: a proteomics perspective

Mass spectrometry-based proteomics and peptidomics for systems biology and biomarker discovery

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