Characterization of the alpha-amylase receptor of Streptococcus gordonii NCTC 7868.
The purpose of the work described here was to investigate the mechanisms involved in the binding of salivary alpha-amylase to Streptococcus gordonii NCTC 7868 (Challis). Of six types of alpha-amylase studied, only mammalian forms of the enzyme were found to bind to S. gordonii cells. Salivary alpha-amylase binding was inhibited by treatment of cells with trypsin and pronase, but not with pepsin or sodium periodate. Presence of starch, dextrin, or maltoheptaose partially inhibited binding of the enzyme to S. gordonii. Both mutanolysin extracts of cells and culture supernatants contained alpha-amylase-binding activity, which was partially purified by Sepharose CL-6B and DEAE-ion-exchange chromatography. Western blotting detected four putative receptor bands--65 kDa, 15 kDa, 12.5 kDa, and one with a very high molecular weight; the lower-molecular-weight components may be products of proteolytic degradation of the high-molecular-weight material, but their true relationship has yet to be determined. Pre-treatment of salivary alpha-amylase with these putative receptors partially inhibited subsequent binding of the enzyme to S. gordonii cells. When bound to cells, only 19% of the salivary alpha-amylase activity was detectable, suggesting that alpha-amylase binds to the receptor at or near the active site of the enzyme.
Douglas
C W
CW
Department of Oral Pathology, School of Clinical Dentistry, University of Sheffield, United Kingdom.
D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D010664:Phenylmethylsulfonyl Fluoride / Q000378:metabolism; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D013291:Streptococcus / Q000378:metabolism*; D000516:alpha-Amylases / Q000737:chemistry / Q000378:metabolism*
D011956:Receptors, Cell Surface; D012471:Salivary Proteins and Peptides; D012967:Sodium Dodecyl Sulfate; D010664:Phenylmethylsulfonyl Fluoride; D000516:alpha-Amylases