High-performance liquid chromatography of amino acids, peptides, and proteins. 123. Dynamics of peptides in reversed-phase high-performance liquid chromatography.
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 beta-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, beta-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.
Purcell
A W
AW
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
Aguilar
M I
MI
Hearn
M T
MT
D000595:Amino Acid Sequence; D001839:Bombesin / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005934:Glucagon / Q000032:analysis* / Q000737:chemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D010649:Phenylalanine / Q000032:analysis* / Q000737:chemistry; D011487:Protein Conformation; D012822:Silicon Dioxide / Q000737:chemistry; D013816:Thermodynamics; D001615:beta-Endorphin / Q000032:analysis* / Q000737:chemistry
D010649:Phenylalanine; D001615:beta-Endorphin; D012822:Silicon Dioxide; D005934:Glucagon; D001839:Bombesin