Intestinal mucins: The binding sites for Salmonella typhimurium

Mucus-bacterial interactions in the gastrointestinal tract and their impact on subsequent enteric infections are poorly delineated. In the present stu dy, we have examined the binding ofSalmonella typhimurium to rat intestinal mucus and characterized a mucus protein (Mucus-Rs) which specifically bind s to S. . Both virulent (1402/84), and avirulent (SF 1835) S. typhimurium w ere observed to bind to crude mucus, however, the virulent strain showed 6 fold more binding as compared to avirulent strain. Fractionation of crude m ucus on sepharose CL-6B resolved it into three major peaks. Maximal bacteri al binding was observed with a high mol. wt. glycoprotein corresponding to neutral mucin. SDS-PAGE of purified protein (termed Mucus-Rs) under non red ucing conditions showed it to be a homogenous glycoprotein (mol. wt. 250 kD a), while under reducing conditions, three bands corresponding to mol. wt. of 118,75 and 60 kDa were observed. Pretreatment of Mucus-Rs with pronase, trypsin and sodium metaperiodate markedly inhibited bacterial binding. GLC analysis of Mucus-Rs showed it to contain Mannose, Glucose, Galactose, Gluc osamine, Galactosamine and Sialic acid as main sugars. Competitive binding in the presence of various sugars and lectins indicated the involvement of mannose in the mucus-bacterial interactions. The Mucus-Rs binding was highl y specific for S. typhimurium; no significant binding was seen with E. coli and V. cholerae. Thus, we conclude that S. typhimurium specifically binds to a 250 kDa neutral mucin of intestinal tract. This binding appears to occ ur via specific adhesin-receptor interactions involving bacterial pili and mannose of neutral mucin.