Ultracentrifugation of Salivary Mucins
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The salivary mucins contribute to the protection of enamel against the attack by bacteria and their metabolic products among others by the formation of a protein pellicle on its surface. To study the interaction of salivary mucins with enamel and bacteria, pure and well characterized mucins should be available. A widely used method for the isolation of salivary mucins is based on clot formation with a quaternary ammonium compound, followed by fractional precipitation of the mucin from the resolved clot by ethanol (1). This method is only convenient if the glycoproteins are rich in sialic acid or sulphate. Another effective method used heating at 100°C to remove contaminating proteins, followed by further purification on a Biogel P-300 column (2,3). However this method could not be applied for the preparation of a mucin from the human sublingual glands. We therefore replaced the Biogel step by ultracentrifugation and purified the human sublingual mucin (HSL), the human submandibular mucin (HSM), the mouse sublingual and submandibular mucins (MSL and MSM) in this way. HSM was studied in more detail, because it has been described (3) that the incorporation of proteolytic digestion and/or -S-S-bond reduction in the purification procedure results in preparations with very low protein values.
KeywordsSialic Acid Migration Rate Clot Formation Quaternary Ammonium Compound Proteolytic Digestion
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