Purification and characterization of an adhesin from Pasteurella haemolytica
Source: Glycobiology, Volume 10, Number 1, 1 JANUARY 2000 , pp. 31-37(7)
Publisher: Oxford University Press
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.haemolyticato 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.
Document Type: Research Article
Affiliations: 1: Departamento de Bioquímica, Instituto Nacional de Enfermedades Respiratorias, Secretaría de Salud, México, 2: Laboratoire de Chimie Biologique, UMR 8576 du CNRS, Université des Sciences et Technologies de Lille, 59655 Villeneuve d’Ascq, France, 3: Departamento de Patología, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, and 4: Departamento de Bioquímica, Facultad de Medicina UNAM, PO Box 70159, 04510 Mexico DF
Publication date: 1 JANUARY 2000
- Established as the leading journal in the field, Glycobiology provides a unique forum dedicated to research into the biological functions of glycans, including glycoproteins, glycolipids, proteoglycans and free oligosaccharides, and on proteins that specifically interact with glycans (including lectins, glycosyltransferases, and glycosidases).