Glycoprofiling of the Human Salivary Proteome

Melissa Sondej,Joseph A Loo,Jona Takashima,Paul C Denny,David T Wong,Yan Si,Yongming Xie,Prasanna Ramachandran,Wenyuan Shi,Patricia A Denny

doi:10.1007/s12014-008-9021-0

Melissa Sondej, Joseph A Loo + Show 8 more

Open Access

https://doi.org/10.1007/s12014-008-9021-0

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Abstract

Glycosylation is important for a number of biological processes and is perhaps the most abundant and complicated of the known post-translational modifications found on proteins. This work combines two-dimensional polyacrylamide gel electrophoresis (2-DE) 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). 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 of the salivary proteins are glycosylated and that the pattern is more widespread than is demonstrated by the glycoprotein stained gel. Finally, the co-reactivity between two lectins was measured to determine the glycan structures that are most and least often associated with one another along with the population variation of the lectin reactivity for 66 individuals.

Highlights

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
Of the 78 additional spots picked, 69 spots yielded a positive identification. When comparing these protein identifications to our previous work with whole saliva that used both 2-D gel electrophoresis and bottom-up LC-mass spectrometry (MS)/MS and MALDITOF MS approaches [8], 50 new proteins were identified in this study
The data were submitted to the database that houses the compiled data from the Human Salivary Proteome Project, representing work from the Scripps Research Institute/University of Rochester, the University of California—Los Angeles/University of Southern California, and the University of California— San Francisco

Summary

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. Clin Proteom (2009) 5:52–68 microbes, epithelial cells, nasal and bronchial secretions, and serum products. These components can provide clues to local and/or systemic diseases and disorders of the human body [1]. Numerous studies have sought to develop a complete catalog of the proteins and peptides found in human saliva [2,3,4,5,6,7,8,9,10,11,12]. Proteins are only one chapter in gaining a comprehensive catalog of all the components in saliva; another is to characterize the post-translational modifications of the proteins and peptides that compose the human salivary catalog

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