Glycoprofiling of the Human Salivary Proteome |
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Authors: | Melissa Sondej Patricia A Denny Yongming Xie Prasanna Ramachandran Yan Si Jona Takashima Wenyuan Shi David T Wong Joseph A Loo Paul C Denny |
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Institution: | 1. Department Chemistry and Biochemistry, University of California at Los Angeles, P. O. Box 951569, Los Angeles, CA, 90095, USA 2. Division of Oral Biology and Department of Medicine and Microbiology, Immunology and Molecular Genetics, University of California at Los Angeles, Los Angeles, CA, USA 3. Division of Diagnostic Sciences, School of Dentistry, University of Southern California, Los Angeles, CA, USA 4. Division of Oral Biology and Dental Research Institute, School of Dentistry, University of California at Los Angeles, Los Angeles, CA, USA 5. Department of Biological Chemistry, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA, USA
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Abstract: | Introduction Glycosylation is an important component for a number of biological processes and is perhaps the most abundant and complicated
of the known post-translational modifications found on proteins.
Methods This work combines two-dimensional (2-D) polyacrylamide gel electrophoresis and lectin blotting to map the salivary glycome
and mass spectrometry to identity the proteins that are associated with the glycome map. A panel of 15 lectins that recognize
six sugar-specific categories was used to visualize the type and extent of glycosylation in saliva from two healthy male individuals.
Lectin blots were compared to 2-D gels stained either with Sypro Ruby (protein stain) or Pro-Q Emerald 488 (glycoprotein stain).
Results Each lectin shows a distinct pattern, even those belonging to the same sugar-specific category. In addition, the glycosylation
profiles generated from the lectin blots show that most salivary proteins are glycosylated and that the profiles are more
widespread than is demonstrated by the glycoprotein-stained gel. Finally, the coreactivity between lectins was measured to
determine what types of glycan structures are associated with one another and also the population variation of the lectin
reactivity for 66 individuals were reported.
Conclusions This starting 2-D gel glycosylation reference map shows that the scientifically accepted, individual oligosaccharide variability
is not limited to a few large glycoproteins such as MUC5B, but are found on most members of the salivary proteome. Finally,
in order to see the full range of oligosaccharide distribution, multiple reagents or lectins are needed.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Glycosylation Human whole saliva Lectin blotting Two-dimensional gel electrophoresis |
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