Purification and characterization of an adhesin from Pasteurella haemolytica

Abstract

We purified an adhesin from Pasteurella. haemolytica by affinity chromatography using glutaraldehyde treated rabbit erythrocytes stroma. The adhesin is a protein of 68 kDa, as determined by SDS-PAGE, and the most abundant amino acids constituting this protein were Gly, Ser, Glx, and Ala, and low concentrations of Cys, Met, and Tyr residues were also found. The N-terminal sequence of the adhesin is ANEVNVYIYKQPYLI. No carbohydrate residues were detected. The adhesin agglutinated rabbit erythrocytes but when the latter were desialylated or pronase treated the agglutinating activity was abolished. The agglutinating activity of the adhesin was inhibited with N-acetyl-D-glucosamine (GlcNAc), and in a lesser degree with N-acetyl-neuraminic acid (NeuAc). GalNAc, N-glycolyl-neuraminic acid, N-deacetylated GlcNAc, or neutral sugars do not modify the activity of the adhesin. The equatorial -OH on C4 and the NH-acetylated group on C2 from GlcNAc, as well as the 4-OH and NH-acetylated group on C5 from NeuAc seem to be responsible for the interaction with the adhesin. The protein is divalent cation-dependent and thermolabile. As for the agglutinating activity, the adhesion of P.haemolytica to tracheal cell-cultures was inhibited by GlcNAc, NeuAc or the purified adhesin, strongly suggesting that the P.haemolytica adhesin plays an important role in infection.

Role ofMannheimia haemolyticaleukotoxin in the pathogenesis of bovine pneumonic pasteurellosis

“…These findings suggest that virulent M. haemolytica ST1 may possess adhesion factors that enable the organism to establish microcolonies in the URT. The interaction of a 68-kDa adhesin from M. haemolytica with rabbit erythrocytes and bovine tracheal epithelial cells was inhibited by N-acetyl-D-glucosamine and, to a lesser degree, by N-acetyl-neuraminic acid (Jaramillo et al, 2000). Furthermore, the finding that pretreatment of tracheal epithelial cells with neuraminidase or protease inhibits binding suggested that the host receptor for M. haemolytica adhesin binding is a sialoglycoprotein (Jaramillo et al, 2000).…”

Sialic acid-specific lectins: occurrence, specificity and function

“…Different members of the Pasteurellaceae group, such as Haemophilus influenzae, Actinobacillus actinomycetemcomitans and Pasteurella haemolytica have been found to possess Sia-specific lectins [68,69]. The HMW1 and HMW2 proteins from H. influenzae are high-molecularweight adhesins that mediate binding to cultured epithelial cells.…”

Identification ofMannheimia haemolyticaAdhesins Involved in Binding to Bovine Bronchial Epithelial Cells

“…A 68-kDa adhesin with hemagglutinating activity for rabbit erythrocytes specifically recognizes glycoproteins containing N-acetyloglucosamine (GlcNAc) on bovine neutrophils and tracheal epithelial cells (14,23). Binding of this 68-kDa adhesin to bovine neutrophils induces an oxidative burst (14).…”