Specific and non-specific affinities of the extracellular glucosyltransferase complex of Streptococcus mutans 6715.
Glucosyltransferases (GTF) from different strains of streptococci exhibited different elution profiles when fractionated on insoluble-dextran affinity columns. The proportions of unadsorbed and adsorbed GTF were not related to their extent of stimulation by exogenous dextran, and GTF preparations exposed to, and freed from, clinical dextran prior to fractionation lost their ability to bind to the dextran columns. Different proportions of bound GTF were released by irrigation of columns with different concentrations of salt and clinical dextran, and the "specific" binding and release of GTF exhibited by a column possessing covalently linked, clinical dextran ligands was duplicated on a control column that did not possess the dextran ligands. These results, and the high affinity of GTF for hydrophobic alkyl (Shaltiel) ligands, demonstrate that ionic and hydrophobic properties of impure GTF aggregates may lead to erroneous characterization of the dextran affinity of some protein fractions. Fractionations on DEAE-Sepharose and on hydroxylapatite showed that the two dextran-dependent GTF activities (GTF-S and GTF-I) were present in the major enzyme fraction (Streptococcus mutans 6715) recovered from a Sephacryl S-200 affinity column. A minor, dextran-independent GTF was not adsorbed onto the Sephacryl column. The presence of SDS (0.005%) and Triton X100 (0.01%) stabilized GTF activity during gel filtration and improved the separation of GTF-S and GTF-I in hydroxylapatite fractionation of the highly aggregated enzyme. A comparable separation of the two enzyme forms on DEAE-Sepharose was achieved only if T10 dextran (10 mg/mL) was included with the detergent mixture in the column irrigant.
D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel; D005632:Fructose / Q000378:metabolism; D005936:Glucans / Q000378:metabolism; D005964:Glucosyltransferases / Q000378:metabolism*; D013295:Streptococcus mutans / Q000201:enzymology*; D013395:Sucrose / Q000378:metabolism
D003911:Dextrans; D005936:Glucans; D005632:Fructose; D013395:Sucrose; D005964:Glucosyltransferases