Peptide mapping of 125I-labelled membrane protein of influenza viruses by reverse-phase high-performance liquid chromatography.
The resolution potential of reverse-phase high-performance liquid chromatography (HPLC) for peptide analysis of hydrophobic viral membranes has been investigated, using as model the membrane (M) protein of influenza virus. Proteolytic digests of 125I-labelled M protein and CNBr fragments, extracted from radioiodinated whole virus, have been separated on a uBondapak C18 column with an isopropanol or acetonitrile solvent system. Peptide mapping of trypsin digests of M protein from A/PR/8/34 (H1N1) and A/chicken/Germany/N/49 (H10N7) viruses was identical, whereas Staphylococcus aureus V8 protease digests showed minor differences in at least two peptides. The results also show that HPLC is a powerful tool for the separation of proteolytic digests of viral proteins, since the peptide maps are highly reproducible and recovery was greater than 85%.
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D009980:Influenza A virus / Q000032:analysis*; D007457:Iodine Radioisotopes; D008565:Membrane Proteins / Q000032:analysis*; D010455:Peptides / Q000032:analysis*; D014764:Viral Proteins / Q000032:analysis*
D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D010455:Peptides; D014764:Viral Proteins