Preparation of gastric mucin "STP-domains" using pronase digestion and chemical deglycosylation.
In this work we present a method of mucin protein backbone fragments preparation, which are rich in serine, threonine and proline amino acid residues. The purified native gastric mucin was reduced, the pronase digested and chemically deglycosylated. The obtained apomucin preparations contained slight quantities of residual carbohydrates (0-1%). In O-glycosylation reaction in vitro, with the participation of GalNAc-transferase, the amounts of incorporated [14C]GalNAc to apomucin fractions were about twofold greater in relation to the deglycosylated non-digested mucin subunits. We also demonstrate the usefulness of immunoblotting technique of apomucin preparations detection.
Paszkiewicz-Gadek
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Department of General and Organic Chemistry, Medical Academy of Białystok.
Porowska
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Gindzieński
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D004591:Electrophoresis, Polyacrylamide Gel; D005752:Gastric Mucins / Q000302:isolation & purification*; D006031:Glycosylation; D006801:Humans; D011402:Pronase
D005752:Gastric Mucins; C053366:apomucin; D011402:Pronase