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KMID : 0829220140380060381
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Korean Journal of Oral and Maxillofacial Pathology
2014 Volume.38 No. 6 p.381 ~ p.388
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High Performance Liquid Chromatography Analysis of Human Salivary Protein Complexes
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Kim Yeon-Sook
Lee Suk-Keun
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Abstract
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Salivary proteins include numerous functional proteins which play important roles not only for the food-intake but also for theprotective and defensive mechanisms. In the present study the compositions of salivary proteins were analyzed by different methods,including electrophoresis and high performance liquid chromatography (HPLC). In hydrophobic protein HPLC analysis the parotid salivagradually produced macromolecular complexes when agitated in refrigerator until 30 minutes. These salivary protein complexes weredigested by neuraminidase, and then migrated more rapidly in native tris glycine gel than the control. Therefore, it was assumed thatthe glycosylated proteins of parotid saliva became gradually aggregated to form salivary protein complexes similar to those of wholesaliva. The salivary protein complexes were easily degenerated in different experimental buffers, i.e., SDS buffer, tris glycine buffer,methanol, etc., and resulted non-specific patterns in electrophoresis and HPLC. Therefore, it was presumed that the salivary proteincomplexes was made by the hydrophobic interaction as well as electrostatic attraction between salivary proteins. These data indicatedthat to know the real pattern of salivary protein complexes in vivo the whole saliva should be analyzed by HPLC using non-adheringcolumn with isoelectric buffer. Consequently, the whole saliva was analyzed by HPLC using reverse phase SuperoseTM column with20 mM potassium phosphate buffer, and two prominent peaks of salivary protein complexes were consistently found in every people.
These salivary protein complex peaks were relatively stable up to 6 hours after saliva collection when the whole saliva was keptin refrigerator during experiment. Therefore, it is suggested that the salivary protein complex patterns are characteristic macromolecularstructures of whole saliva, which are also applicable as a diagnostic point in different saliva-associated diseases.
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KEYWORD
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Human saliva, Salivary Protein Complexes, HPLC
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