Mass Spectrometric Characterization of the Glycosylation Pattern of HIV-gp120 Expressed in CHO Cells
Citations Over TimeTop 11% of 2000 papers
Abstract
An analytical approach is reported for the characterization of the specific glycans found on highly glycosylated proteins based on a combination of specific proteolysis and deglycosylation combined with two different mass spectrometric approaches, matrix-assisted laser desorption/ionization mass spectrometry, and nanoelectrospray mass spectrometry/tandem mass spectrometry using a hybrid quadrupole-time-of-flight tandem mass spectrometer. The high resolution and mass accuracy of the mass spectrometric data obtained on the hybrid instrument combined with the high parent mass capabilities are shown to be extremely useful in the site-specific assignment of heterogeneous glycans. Using this methodology, 25 of 26 consensus glycosylation sites on HIV-1(SF2) gp120, expressed in Chinese hamster ovary cells, could be assigned. Good correlations between the relative abundances of members of heterogeneous series in the matrix-assisted laser desorption/ionization mass spectra and the nanoelectrospray mass spectra were observed, indicating that the mass spectrometric data reflected the actual abundances of the members of the series. These data were incorporated with molecular modeling based on the solved structure of a mutant truncated, highly deglycosylated gp120 to propose a structural model for the completely glycosylated form.
Related Papers
- → Assignment of the mass number to peaks in high-resolution mass spectra acquired via the counting of ions(1988)5 cited
- → Analysis of Mixed Drugs by Mass Spectrometry. II(1965)5 cited
- → Mass spectra of some steroidal diols(1972)6 cited
- → Anomalous peaks in mass spectra; effects of pre-analysis dissociation and charge exchange events(2002)2 cited
- → Studies on the mass spectrometry of natural products: V. Negative ion fast atom bombardment and collisional activation mass spectrometry in the structural analysis of oligoglycosides(1987)