Glycoprotein profiling by electrospray mass spectrometry

This work compares several different methods of site-specific analysis of glycoproteins using electrospray mass spectrometry. The glycoprotein, oLHα (ovine luteinizing hormone, α-subunit) was chosen as an appropriate example protein for these studies because of its biological relevance and extreme m...

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Bibliographic Details
Published in:Journal of the American Society for Mass Spectrometry 2004-05, Vol.15 (5), p.750-758
Main Authors: Jiang, Hui, Desaire, Heather, Butnev, Vladimir Y, Bousfield, George R
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Binding
Biological and medical sciences
Carbohydrates
Data acquisition
Electrospraying
Fundamental and applied biological sciences. Psychology
Glycoproteins
Glycoproteins - chemistry
Glycoproteins - isolation & purification
Ions
Mass spectrometry
Monosaccharides - analysis
Monosaccharides - chemistry
Peptide Fragments - chemistry
Proteins
Quadrupoles
Reproducibility of Results
Scientific imaging
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization - methods
Spectroscopy
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Summary:This work compares several different methods of site-specific analysis of glycoproteins using electrospray mass spectrometry. The glycoprotein, oLHα (ovine luteinizing hormone, α-subunit) was chosen as an appropriate example protein for these studies because of its biological relevance and extreme microheterogeneity. More than 20 unique glycoforms were detected for this glycoprotein at the Asn 56 site of oLHα. The carbohydrates present at this site affect receptor binding affinity, so understanding the great variety in the composition of these carbohydrates is important in studying ligand binding interactions. MS data was acquired on a quadrupole ion trap, a triple quadrupole, and a quadrupole time of flight mass spectrometer, and carbohydrate composition at the Asn 56 site of oLHα was determined using these instruments. Additionally, neutral loss and precursor ion scanning modes were also used to identify the glycoforms present, and these techniques were compared to the standard MS data. Of the three instruments compared in the study, the qTOF mass spectrometer achieved the lowest sample consumption, but all three instruments were useful in profiling the glycopeptide composition.
ISSN:1044-0305
1879-1123
DOI:10.1016/j.jasms.2004.01.009