Highly Resolutive Separations of Hardly Soluble Synthetic Polypeptides by Capillary Electrophoresis

Miramon, Hélène; Cavelier, Florine; Martinez, Jean; Cottet, Hervé

doi:10.1021/ac902211f

Cited by 14 publications

(14 citation statements)

References 44 publications

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“…PCR was performed on an ABI Veriti thermal cycler (Applied Biosystems, Grand Island, NY). CGG repeat numbers were determined by agarose gel electrophoresis of PCR products and confirmed by capillary electrophoresis on an ABI 3730xl DNA Analyzer with POP-7 195 polymer using a 50-cm array [40], which is capable of defining exact CGG repeat number on samples with 200 CGG repeats or less.…”

Section: Methodsmentioning

confidence: 99%

CGG Repeats in the 5’UTR of FMR1 RNA Regulate Translation of Other RNAs Localized in the Same RNA Granules

Rovozzo¹,

Korza²,

Baker³

et al. 2016

PLoS ONE

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CGG repeats in the 5’UTR of Fragile X Mental Retardation 1 (FMR1) RNA mediate RNA localization and translation in granules. Large expansions of CGG repeats (> 200 repeats) in FMR1, referred to as full mutations, are associated with fragile X syndrome (FXS). Smaller expansions (55–200 repeats), referred to as premutations, are associated with fragile X tremor ataxia syndrome (FXTAS) and fragile X premature ovarian insufficiency (FXPOI). TMPyP4 is a porphyrin ring compound that destabilizes CGG repeat RNA secondary structure. Here we show that exogenous CGG repeat RNA by itself, lacking the FMRP ORF, microinjected into hippocampal neurons is localized in RNA granules and inhibits translation of ARC RNA, which is localized in the same granules. TMPyP4 rescues translation of ARC RNA in granules. We also show that in human premutation fibroblasts with endogenous CGG repeat expansions in the FMR1 gene, translation of ARC RNA is inhibited and calcium homeostasis is disrupted and both phenotypes are rescued by TMPyP4. Inhibition of granule translation by expanded CGG repeats and rescue of granule translation by TMPy4, represent potential pathogenic mechanism and therapeutic strategy, respectively, for FXTAS and FXPOI.

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Section: Methodsmentioning

confidence: 99%

CGG Repeats in the 5’UTR of FMR1 RNA Regulate Translation of Other RNAs Localized in the Same RNA Granules

Rovozzo¹,

Korza²,

Baker³

et al. 2016

PLoS ONE

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“…The fourth category is the separation of polymers bearing a single charge or no charge. For the latter category, the reader is referred to the pioneering work of the groups of Cottet [25,26] and Cifuentes [24,27].…”

Section: Free Solution Capillary Electrophoresis (Ce)mentioning

confidence: 99%

Separation and Characterization of Synthetic Polyelectrolytes and Polysaccharides with Capillary Electrophoresis

Thevarajah

Gaborieau

Castignolles

2014

Advances in Chemistry

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The development of macromolecular engineering and the need for renewable and sustainable polymer sources make polymeric materials progressively more sophisticated but also increasingly complex to characterize. Size-exclusion chromatography (SEC or GPC) has a monopoly in the separation and characterization of polymers, but it faces a number of proven, though regularly ignored, limitations for the characterization of a number of complex samples such as polyelectrolytes and polysaccharides. Free solution capillary electrophoresis (CE), or capillary zone electrophoresis, allows usually more robust separations than SEC due to the absence of a stationary phase. It is, for example, not necessary to filter the samples for analysis with CE. CE is mostly limited to polymers that are charged or can be charged, but in the case of polyelectrolytes it has similarities with liquid chromatography in the critical conditions: it does not separate a charged homopolymer by molar mass. It can thus characterize the topology of a branched polymer, such as poly(acrylic acid), or the purity or composition of copolymers, either natural ones such as pectin, chitosan, and gellan gum or synthetic ones.

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“…Methanolic BGEs composed of 50 mM Tris, 100 mM chloroacetic acid, pH MeoH 7.8, were applied for investigation of complexes of water insoluble valinomycin (macrocyclic dodecadepsipeptide antibiotic) with alkali metal ions 101–103. Mixed hydro‐organic hexafluoroisopropanol/water (80/20, v/v) solvent and BGE composed of 0.4 M formic acid and containing 1.0% m/v hydroxyethyl cellulose as an adsorption reducing agent enabled highly resolutive separation of in‐water hardly soluble synthetic polyalanines up to 110 residues 104, see Fig. 3.…”

Section: Separations In Different Ce and Cec Modesmentioning

confidence: 99%

Recent developments in CE and CEC of peptides (2009–2011)

Kašička

2011

Electrophoresis

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The review brings a comprehensive survey of the recent developments of high-performance electroseparation methods in capillary and microchip formats: zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography and electrochromatography. Applications of these techniques to analysis, isolation, purification and physicochemical and biochemical characterization of peptides are described. Advances in the investigation of electromigration properties of peptides, and in the methodology of their analysis, such as sample preparation, adsorption suppression, EOF control and detection, are presented. New developments, in particular, CE and CEC modes are reported and several types of their applications to peptide analysis are described: conventional qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence and chiral analysis, and peptide mapping of proteins. Some micropreparative peptide separations are shown and capabilities of CE and CEC techniques to provide relevant physicochemical characteristics of peptides are demonstrated.

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Highly Resolutive Separations of Hardly Soluble Synthetic Polypeptides by Capillary Electrophoresis

Cited by 14 publications

References 44 publications

CGG Repeats in the 5’UTR of FMR1 RNA Regulate Translation of Other RNAs Localized in the Same RNA Granules

CGG Repeats in the 5’UTR of FMR1 RNA Regulate Translation of Other RNAs Localized in the Same RNA Granules

Separation and Characterization of Synthetic Polyelectrolytes and Polysaccharides with Capillary Electrophoresis

Recent developments in CE and CEC of peptides (2009–2011)

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