Isoelectric Point Separations of Peptides and Proteins

Pergande, Melissa R; Cologna, Stephanie M.

doi:10.3390/proteomes5010004

Cited by 79 publications

(53 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Basic aspects of IEF including CIEF of peptides and proteins, such as choice of carrier ampholytes, p I markers, sample application, and mobilization, avoiding of isoelectric precipitation, immobilized pH gradient, and selected applications are in detail described and discussed in the tutorial and review articles . Latest developments in CIEF coupled to MS detection are presented in the recent review whereas the present state of art in microchip IEF and its applications are described in .…”

Section: Separations By the Particular Ce And Cec Methodsmentioning

confidence: 99%

“…Useful information and practical advices for CZE and CIEF analysis of peptides and proteins can be found also in the tutorial and in the review articles summarizing recent applications of multivariate techniques for optimization of CE methods and dealing with (un)suitability of pH buffers in biological, biochemical, and environmental studies, and providing data on their interactions with metal ions . Important parameter for selection of cationic or anionic mode of CE separation of peptides is their isoelectric point (p I ), which can be found in the database , or calculated or experimentally determined by IEF in capillary or slab gel format .…”

Section: Electromigration Properties Of Peptides and Selection Of Sepmentioning

confidence: 99%

“…Nevertheless, due to the approximative character of these models, only approximative values of charge and size (Stokes radius or M r ) are obtained from these measurements. Another important peptide/protein parameter related to charge, p I , can be determined by CIEF or by CZE , but also in this case one should take into account that the obtained p I values are dependent on the composition of carrier ampholytes in CIEF or BGEs in CZE, and other experimental conditions used.…”

Section: Applicationsmentioning

confidence: 99%

See 2 more Smart Citations

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Kašička

2017

Electrophoresis

View full text Add to dashboard Cite

The review brings a comprehensive overview of recent developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) to analysis, microscale isolation, purification, and physicochemical and biochemical characterization of peptides in the years 2015, 2016, and ca. up to the middle of 2017. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis (sample preseparation, preconcentration and derivatization, adsorption suppression and EOF control, and detection) are described. New developments in particular CE and CEC methods are presented and several types of their applications to peptide analysis are reported: 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 methods to provide important physicochemical characteristics of peptides are demonstrated.

show abstract

Section: Separations By the Particular Ce And Cec Methodsmentioning

confidence: 99%

Section: Electromigration Properties Of Peptides and Selection Of Sepmentioning

confidence: 99%

Section: Applicationsmentioning

confidence: 99%

See 1 more Smart Citation

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Kašička

2017

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Out of 20 fractions of G. sylvestre extract obtained from Rotofor IEF, the dark-colored molecules (terpenoid saponins) were migrated and enriched at the anode end (pH 2-3) and the light-yellow clear fractions were observed at cathode end (pH 8-9) ( Figure 2). Aliquots (20 l) from each fraction (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) were separated after reducing and boiling on 15% SDS-PAGE. The Coomassie bluestained gel shows the diffused polypeptide band of about 5 kDa that is enriched in fractions 16-19 ( Figure 3).…”

Section: Resultsmentioning

confidence: 99%

Separation of Bioactive Small Molecules, Peptide from Natural Products and Proteins from Pathogenic Microbe by Rotofor-based Preparative Isoelectric Focusing (IEF) Method

Veerapandian

Paudyal

Chang

et al. 2019

Preprint

View full text Add to dashboard Cite

The primary purpose of purifying the bioactive small molecules is obtaining enough amount of specific molecules for their downstream applications. There are various methods available for serving this purpose. However, not all can afford for cost-effective purification methods, especially for complex biological samples. Here we demonstrate, the Rotofor based preparative IEF separation which shows the high possibility of separating and enriching desired molecules, including small molecules and peptides from the complex plant extract. This method is wellsuited for use at any stage of a purification scheme and has been used for complex biological samples quantification and characterization. As a proof of concept, we fractionated Gymnema sylvestre plant extract and isolated a family of terpenoid saponin molecules at the acidic end and small peptide molecule near the basic end. To our knowledge, this is a first report that Rotofor can be used to fractionate bioactive small molecules from natural product sources. We also showed effective microbial protein separation using Candida albicans fungus as a model system. The Rotofor based IEF has appealing features such as low cost, simplicity, high application potential and can be performed in any laboratory with the instrument setup.

show abstract

“…The transcription of genes encoding HSPs is controlled by regulatory proteins called HSFs (Hu et al 2009). 2-D technique is used to separate ampholytic components, improving protein recognition, and to analyze and compare synthesis, turnover, and modification of many proteins during development or in response to environmental changes (Pergande andCologna 2017, Schuster andDavies 1983). Chen et al (2014) used 2-DE to identify combined drought-and heat-responsive protein spots in maize leaves, which found about 450 spots were reproducibly detected on each gel.…”

Section: Discussionmentioning

confidence: 99%

Proteomic analysis of heat shock proteins in maize (Zea mays L.)

et al. 2019

View full text Add to dashboard Cite

Background: Maize is one of the important cereal food crops in the world. High temperature stress causes adverse influence on plant growth. When plants are exposed to high temperatures, they produce heat shock proteins (HSPs), which may impart a generalized role in tolerance to heat stress. Proteome analysis was performed in plant to assess the changes in protein types and their expression levels under abiotic stress. The purpose of the study is to explore which proteins are involved in the response of the maize plant to heat shock treatment. Results:We investigated the responses of abundant proteins of maize leaves, in an Egyptian inbred line of maize "K1", upon heat stress through two-dimensional electrophoresis (2-DE) on samples of maize leaf proteome. 2-DE technique was used to recognize heat-responsive protein spots using Coomassie Brilliant Blue (CBB) and silver staining. In 2-D analysis of proteins from plants treated at 45°C for 2 h, the results manifested 59 protein spots (4.3%) which were reproducibly detected as new spots where did not present in the control. In 2D for treated plants for 4 h, 104 protein spots (7.7%) were expressed only under heat stress. Quantification of spot intensities derived from heat treatment showed that twenty protein spots revealed clear differences between the control and the two heat treatments. Nine spots appeared with more intensity after heat treatments than the control, while four spots appeared only after heat treatments. Five spots were clearly induced after heat treatment either at 2 h or 4 h and were chosen for more analysis by LC-MSMS. They were identified as ATPase beta subunit, HSP26, HSP16.9, and unknown HSP/Chaperonin. Conclusion:The results revealed that the expressive level of the four heat shock proteins that were detected in this study plays important roles to avoid heat stress in maize plants.

show abstract

Isoelectric Point Separations of Peptides and Proteins

Cited by 79 publications

References 87 publications

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Separation of Bioactive Small Molecules, Peptide from Natural Products and Proteins from Pathogenic Microbe by Rotofor-based Preparative Isoelectric Focusing (IEF) Method

Proteomic analysis of heat shock proteins in maize (Zea mays L.)

Contact Info

Product

Resources

About