Purification of the receptor for the N-acetyl-D-glucosamine specific adhesin of Mannheimia haemolytica from bovine neutrophils.
The GlcNAc-specific adhesin from Mannheimia haemolytica (MhA) has been shown to participate in pathogenicity of mannheimiosis due to its capacity to adhere to tracheal epithelial cells and activate the oxidative burst of bovine neutrophils. In this work, we purified the MhA receptor from bovine neutrophils (MhAr) by affinity chromatography on MhA-Sepharose. The MhAr, which corresponded to approximately 2% of the protein from cell lysate, is a glycoprotein mainly composed of Glu, Ala, Ser, Gly, and Asp, without cysteine. The glycan portion, which corresponds to 20% by weight, is composed of GalNAc, GlcNAc, Man, Gal, and NeuAc. The receptor is a 165-kDa glycoprotein, as determined by molecular sieve chromatography under native conditions; SDS-PAGE analysis shows a heterodimer of 83 and 80 kDa subunits. This work suggests that the GlcNAc-containing receptor plays a relevant role by activating bovine neutrophils through non-opsonic mechanisms.
De la Mora
Alfonso
A
Laboratorio de Patología, Instituto de Investigaciones en Ciencias Veterinarias, Universidad Autónoma de Baja California, Mexicali, BC, Mexico.
Suárez-Güemes
Francisco
F
Trigo
Francisco
F
Gorocica
Patricia
P
Solórzano
Carlos
C
Slomianny
Marie-Christine
MC
Agundis
Concepción
C
Pereyra
M Ali
MA
Zenteno
Edgar
E
D000117:Acetylglucosamine / Q000276:immunology / Q000378:metabolism*; D018829:Adhesins, Bacterial / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D016978:Mannheimia haemolytica / Q000276:immunology*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology*; D011971:Receptors, Immunologic / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D016897:Respiratory Burst
D018829:Adhesins, Bacterial; D006023:Glycoproteins; D011971:Receptors, Immunologic; C047105:bacterial adhesin receptor; D000117:Acetylglucosamine