Simulation and optimization of peptide separation by capillary electrophoresis

Cifuentes, Alejandro; Poppe, H.

doi:10.1016/0021-9673(94)80083-9

Cited by 82 publications

(99 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cifuentes and Poppe [18] reported the modification of Grossman's classical linear model, where the logarithmic dependence of mobility on the charge expressed electrostatic charge suppression in highly charged peptides:…”

Section: Models Of Correlations Between Mobility Of Peptides and Theimentioning

confidence: 99%

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

et al. 2004

View full text Add to dashboard Cite

Capillary zone electrophoresis (CZE) has been applied to qualitative analysis, separation, and physicochemical characterization of synthetic insect oostatic peptides (IOPs) and their derivatives and fragments. Series of homologous IOPs were separated in three acidic background electrolytes (BGEs; pH 2.25, 2.30, 2.40) and an alkaline BGE (pH 8.1). Best separation was achieved in acid BGE composed of 100 mM H3PO4, 50 mM Tris, pH 2.25. The effective electrophoretic mobilities, mu(ep), of all IOPs in four BGEs were determined and several semiempirical models correlating effective mobility with charge-to-size ratio (mu(ep) versus q/Mr k) were tested to describe the migration behavior of IOP in CZE. None of models was found to be unambiguously applicable for the whole set of 20 IOPs differing in size (dipeptide - decapeptide) and charge (-2 to +0.77 elementary charges). However, a high coefficient of correlation, 0.9993, was found for the subset of homologous series of IOPs with decreasing number of proline residues at C-terminus, H-Tyr-Asp-Pro-Ala-Prox-OH, x = 6 - 0, for the dependence of mu(ep) on q/Mr k with k = 0.5 for IOPs as anions in alkaline BGE and with k = 2/3 for IOPs as cations in optimized acidic Tris-phosphate BGE. From these dependences the probable structure of IOPs in solution could be predicted.

show abstract

Section: Models Of Correlations Between Mobility Of Peptides and Theimentioning

confidence: 99%

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

et al. 2004

View full text Add to dashboard Cite

show abstract

“…The SPPM used to simulate the peptide migration in CE has been described elsewhere [28]. This program includes a computer program for calculating peptide pK a values, an equation that relates the peptide sequence to its electrophoretic mobility [28], and a coupled program for the prediction of electropherograms [37].…”

Section: Computer Programsmentioning

confidence: 99%

“…A solution that can help to overcome these limitations is the development of theoretical models [28][29][30][31][32][33][34][35] able to predict the CE migration time and peak shape of the peptide in different pH and buffers. Following this idea, a system for the prediction of peptide migration (SPPM) for CE-MS was tested in a previous paper [36].…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis‐mass spectrometry of peptides from enzymatic protein hydrolysis: Simulation and optimization

Simó

Cifuentes

2003

Electrophoresis

Self Cite

View full text Add to dashboard Cite

Two important limitations still exist for the characterization of protein digests by capillary electrophoresis-mass spectrometry (CE-MS): (i) the buffer choice (i.e., the buffer must provide an adequate CE separation without ruining the MS signal), and (ii) the frequent generation of "unexpected" peptidic fragments during the enzymatic protein hydrolysis. In this work, a new approach is used to solve these difficulties, namely a theoretical model that relates the electrophoretic behavior of peptides to their sequence. The effectiveness of this procedure is demonstrated by the fast attainment of good CE-MS conditions for analyzing the peptides obtained from an enzymatic protein hydrolysate in a single run. This strategy can provide useful information for helping to characterize "unexpected" fragments from protein digests.

show abstract

“…In addition to the accurate mass measurement and distribution of peptides in the protein sequence, containing information on the non-random behaviour of trypsin, protein structure, post-translational modifications and database sequence errors, there is also information on the physiochemical properties of the individual peptides. This is primarily hydrophobicity in reversed-phase chromatography [124][125][126][127][128] and size and charge in capillary zone electrophoresis [129][130][131][132][133][134][135][136][137][138]. A predictor of retention time according to Eq.…”

Section: Figure 23 (Color Panel Opposite Side) Lc-fticr (A) and Ce-mentioning

confidence: 99%

“…The chromatographic retention or electrophoretic migration time contains information on the physiochemical properties of the peptides, such as hydrophobicity in RPC and size and charge in standard capillary zone electrophoresis (CZE). A model of chromatographic retention [124][125][126][127][128] or electrophoretic migration [129][130][131][132][133][134][135][136][137][138] can be fitted to experimental data from known proteins. These models are then used to predict the retention time for candidate peptides in a database.…”

Section: Mass Spectrometry Peptide Mass Fingerprinting and Informationmentioning

confidence: 99%

Identification and Characterization of Peptides and Proteins Using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Palmblad

Bergquist

2003

Journal of Chromatography Library

View full text Add to dashboard Cite

Mass spectrometry has in recent years been established as the standard method for protein identification and characterization in proteomics with excellent intrinsic sensitivity and specificity. Fourier transform ion cyclotron resonance is the mass spectrometric technique that provides the highest resolving power and mass accuracy, increasing the amount of information that can be obtained from complex samples. This thesis concerns how useful information on proteins of interest can be extracted from mass spectrometric data on different levels of protein structure and how to obtain this data experimentally. It was shown that it is possible to analyze complex mixtures of protein tryptic digests by direct infusion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry and identify abundant proteins by peptide mass fingerprinting. Coupling on-line methods such as liquid chromatography and capillary electrophoresis increased the number of proteins that could be identified in human body fluids. Protein identification was also improved by novel statistical methods utilizing prediction of chromatographic behavior and the non-randomness of enzymatic digestion. To identify proteins by short sequence tags, electron capture dissociation was implemented, improved and finally coupled on-line to liquid chromatography for the first time. The combined techniques can be used to sequence large proteins de novo or to localize and characterize any labile post-translational modification. New computer algorithms for the automated analysis of isotope exchange mass spectra were developed to facilitate the study of protein structural dynamics. The non-covalent interaction between HIV-inhibitory peptides and the oligomerization of amyloid β-peptides were investigated, reporting several new findings with possible relevance for development of anti-HIV drug therapies and understanding of fundamental mechanisms in Alzheimer's disease.

show abstract

Simulation and optimization of peptide separation by capillary electrophoresis

Cited by 82 publications

References 36 publications

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

Capillary electrophoresis‐mass spectrometry of peptides from enzymatic protein hydrolysis: Simulation and optimization

Identification and Characterization of Peptides and Proteins Using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Contact Info

Product

Resources

About