High-Performance Liquid Chromatography of Amino Acids, Peptides, and Proteins. 123. Dynamics of Peptides in Reversed-Phase High-Performance Liquid Chromatography

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Abstract

The dynamics of several peptides in reversed-phase high-performance liquid chromatography (RP-HPLC) have been investigated on both n-octadecyl (C18) silica and n-butyl (C4) silica sorbents. In particular, the conformational interconversions and the relative rates of chromatographic relaxation of bombesin, glucagon, and β-endorphin on both C18 and C4 n-alkylsilicas were monitored by examining changes in the experimental bandwidths of these peptides as a function of temperature and column residence time under linear gradient elution RP-HPLC conditions. The observed band-broadening trends were correlated with previously derived retention parameters and thermodynamic descriptors of the association process determined for bombesin, β-endorphin, glucagon, and a control peptide, penta-l-phenylalanine. This study confirms that bandwidth measurements can be used as an integral experimental component to study the effect of the secondary structure of peptidic solutes on their RP-HPLC retention behavior. Further, the data demonstrate the utility of RP-HPLC as a tool to examine peptide conformational dynamics at hydrophobic surfaces. The relevance of these results to the general phenomenon of peptide-lipid interactions is discussed in terms of the associated evidence for lipid-induced changes in the conformation of these three bioactive peptides.

Original languageEnglish
Pages (from-to)3038-3047
Number of pages10
JournalAnalytical Chemistry
Volume65
Issue number21
DOIs
Publication statusPublished - 1 Nov 1993

Cite this

@article{8b532953ae5a48308d624a94aae18f7e,
title = "High-Performance Liquid Chromatography of Amino Acids, Peptides, and Proteins. 123. Dynamics of Peptides in Reversed-Phase High-Performance Liquid Chromatography",
abstract = "The dynamics of several peptides in reversed-phase high-performance liquid chromatography (RP-HPLC) have been investigated on both n-octadecyl (C18) silica and n-butyl (C4) silica sorbents. In particular, the conformational interconversions and the relative rates of chromatographic relaxation of bombesin, glucagon, and β-endorphin on both C18 and C4 n-alkylsilicas were monitored by examining changes in the experimental bandwidths of these peptides as a function of temperature and column residence time under linear gradient elution RP-HPLC conditions. The observed band-broadening trends were correlated with previously derived retention parameters and thermodynamic descriptors of the association process determined for bombesin, β-endorphin, glucagon, and a control peptide, penta-l-phenylalanine. This study confirms that bandwidth measurements can be used as an integral experimental component to study the effect of the secondary structure of peptidic solutes on their RP-HPLC retention behavior. Further, the data demonstrate the utility of RP-HPLC as a tool to examine peptide conformational dynamics at hydrophobic surfaces. The relevance of these results to the general phenomenon of peptide-lipid interactions is discussed in terms of the associated evidence for lipid-induced changes in the conformation of these three bioactive peptides.",
author = "Purcell, {Anthony W.} and Aguilar, {Marie Isabel} and Milton Hearn",
year = "1993",
month = "11",
day = "1",
doi = "10.1021/ac00069a016",
language = "English",
volume = "65",
pages = "3038--3047",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
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TY - JOUR

T1 - High-Performance Liquid Chromatography of Amino Acids, Peptides, and Proteins. 123. Dynamics of Peptides in Reversed-Phase High-Performance Liquid Chromatography

AU - Purcell, Anthony W.

AU - Aguilar, Marie Isabel

AU - Hearn, Milton

PY - 1993/11/1

Y1 - 1993/11/1

N2 - The dynamics of several peptides in reversed-phase high-performance liquid chromatography (RP-HPLC) have been investigated on both n-octadecyl (C18) silica and n-butyl (C4) silica sorbents. In particular, the conformational interconversions and the relative rates of chromatographic relaxation of bombesin, glucagon, and β-endorphin on both C18 and C4 n-alkylsilicas were monitored by examining changes in the experimental bandwidths of these peptides as a function of temperature and column residence time under linear gradient elution RP-HPLC conditions. The observed band-broadening trends were correlated with previously derived retention parameters and thermodynamic descriptors of the association process determined for bombesin, β-endorphin, glucagon, and a control peptide, penta-l-phenylalanine. This study confirms that bandwidth measurements can be used as an integral experimental component to study the effect of the secondary structure of peptidic solutes on their RP-HPLC retention behavior. Further, the data demonstrate the utility of RP-HPLC as a tool to examine peptide conformational dynamics at hydrophobic surfaces. The relevance of these results to the general phenomenon of peptide-lipid interactions is discussed in terms of the associated evidence for lipid-induced changes in the conformation of these three bioactive peptides.

AB - The dynamics of several peptides in reversed-phase high-performance liquid chromatography (RP-HPLC) have been investigated on both n-octadecyl (C18) silica and n-butyl (C4) silica sorbents. In particular, the conformational interconversions and the relative rates of chromatographic relaxation of bombesin, glucagon, and β-endorphin on both C18 and C4 n-alkylsilicas were monitored by examining changes in the experimental bandwidths of these peptides as a function of temperature and column residence time under linear gradient elution RP-HPLC conditions. The observed band-broadening trends were correlated with previously derived retention parameters and thermodynamic descriptors of the association process determined for bombesin, β-endorphin, glucagon, and a control peptide, penta-l-phenylalanine. This study confirms that bandwidth measurements can be used as an integral experimental component to study the effect of the secondary structure of peptidic solutes on their RP-HPLC retention behavior. Further, the data demonstrate the utility of RP-HPLC as a tool to examine peptide conformational dynamics at hydrophobic surfaces. The relevance of these results to the general phenomenon of peptide-lipid interactions is discussed in terms of the associated evidence for lipid-induced changes in the conformation of these three bioactive peptides.

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U2 - 10.1021/ac00069a016

DO - 10.1021/ac00069a016

M3 - Article

VL - 65

SP - 3038

EP - 3047

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 21

ER -