Chromatographic characterization of HSV-1 gD 268-284 and IL-6 179-185 synthetic oligopeptides by reversed-phase high-performance liquid chromatography, automated Edman degradation and mass spectrometric analysis.
Two groups of synthetic oligopeptides (namino acid = 7 and 17) were prepared by solid-phase peptide synthesis using the Boc-polystyrene strategy. After deprotection, cleavage and gel permeation, the crude products were analysed by conventional RP-HPLC methods. Separation and isolation of major components were performed on a semi-preparative RP-HPLC column. In order to clarify the primary structure of these products, amino acid analysis, Edman degradation sequence determination and analytical RP-HPLC characterization were applied. The isolated fractions were further assessed by direct molar mass investigation utilizing the fast atom bombardment and 252Cf plasma desorption mass spectrometry. The results with an interleukin-6 oligopeptide corresponding to the 179LRALRQM185 sequence indicate that the single peak product obtained by RP-HPLC separation contains only one component, as verified by amino acid analysis and mass spectrometry. In contrast, the analysis of 268LAPEDPEDSALLEDPVG284-NH2 from HSV-1 gD protein suggests that this large peptide amide showing a single peak after repeated purification by RP-HPLC contains microheterogeneities as revealed by mass spectrometry and sequencing, but not by amino acid analysis.
Bösze
S
S
Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Eötvös L. University, Budapest.
Mák
M
M
Medzihradszky-Schweiger
H
H
Hudecz
F
F
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015850:Interleukin-6 / Q000032:analysis*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis*; D018139:Simplexvirus / Q000737:chemistry; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000379:methods*; D014759:Viral Envelope Proteins / Q000032:analysis*
D015850:Interleukin-6; D009842:Oligopeptides; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1