Analysis of protein digests by capillary high-performance liquid chromatography and on-line fast atom bombardment mass spectrometry.
An HPLC system incorporating a packed capillary C18 column has been utilized for high sensitivity peptide mapping and preparative collection for protein sequencing. This system combined with a Frit-FAB mass spectrometer interface also provides the ability to obtain molecular ions for peptides of enzymatically digested proteins in the time it takes to obtain an HPLC chromatogram. The low flow rates permit introduction of the entire column effluent into the mass spectrometer. Detection limits of 0.5-5 pmol are routine. Proteolytic digests of recombinant human methionyl growth hormone and protein carboxyl methyltransferase have been used to demonstrate the HPLC and mass spectrometer performance.
Department of Protein Chemistry, Genentech, Inc., South San Francisco, California 94080.
W J
WJ
Henzel
J H
JH
Bourell
J T
JT
Stults
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D013006:Growth Hormone / Q000031:analogs & derivatives / Q000032:analysis; D019382:Human Growth Hormone; D006801:Humans; D006868:Hydrolysis; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011497:Protein O-Methyltransferase / Q000032:analysis; D011506:Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D011506:Proteins; D019382:Human Growth Hormone; D013006:Growth Hormone; D011497:Protein O-Methyltransferase