Open AccessThe identification of oral microbial lectins by cell affinity chromatography
Author(s) -
Murray P.A.,
Materese V.,
Hoover C.I.,
Winkler J.R.
Publication year - 1987
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1987.tb01994.x
Subject(s) - fetuin , affinity chromatography , actinomyces naeslundii , biochemistry , fusobacterium nucleatum , sepharose , lectin , chromatography , sialic acid , chemistry , bacteria , sialidase , bacterial cell structure , actinomyces , microbiology and biotechnology , biology , glycoprotein , porphyromonas gingivalis , neuraminidase , enzyme , genetics
Whole‐cell affinity chromatography was used as a novel screening technique for identifying and characterizing oral microbial lectins. First, affinity columns bearing oligosaccharides of defined structure were synthesized as lectin‐binding reagents. Fetuin, transferrin (containing terminal NeuAc residues), asialofetuin and asialotransferrin (with terminal Gal residues) were covalently coupled to Sepharose 6MB and incubated with 3 H‐labeled bacterial suspensions in columns fitted with an 80‐μm nylon filter. Bacteria specifically bound were then eluted with the appropriate sugar (NeuAc or Gal). Fusobacterium nucleatum was the most significant binder, with 80% specifically eluted from the asialo‐derivatives. Actinomyces viscosus and Actinomyces naeslundii also showed unique specificity for galactose. In contrast, Streptococcus sanguis bound in greatest numbers to fetuin, consistent with the presence of a sialic acid‐binding site on these bacteria.