N-Acetylneuraminic acid uptake in Pasteurella (Mannheimia) haemolytica A2 occurs by an inducible and specific transport system.
The N-acetylneuraminic acid (NeuAc) transport system of Pasteurella (Mannheimia) haemolytica A2 was studied when this bacterium was grown in both complex and chemically defined media. Kinetic measurements were carried out at 37 degrees C in 50 mM Tris-HCl buffer, pH 8.0, containing 50 microg/ml bovine serum albumin. Under these conditions, the uptake rate was linear for at least 3 min and the calculated K(m) for NeuAc was 0.1 microM. The transport rate was increased by the addition of several cations and was inhibited by sodium arsenite (95%), N,N'-dicyclohexyl-carbodiimide (50%), and 2,4-dinitrophenol (40%) at final concentration of 1 mM (each). These results support the notion that NeuAc uptake is an active sugar cation symporter. Study of specificities showed that glucosamine, mannose and mannosamine inhibited the transport of NeuAc in this bacterium. Analysis of expression revealed that the NeuAc transport system was induced by NeuAc and by the simultaneous presence of glucose and galactose in the growth medium.
Solana
S
S
Departamento de Bioquímica y Biología Molecular, Universidad de León, Campus de Vegazana, 24007, León, Spain.
Reglero A
A
A
Martínez-Blanco
H
H
Revilla-Nuin
B
B
Bravo
I G
IG
Rodríguez-Aparicio
L B
LB
Ferrero
M A
MA
D019297:2,4-Dinitrophenol / Q000494:pharmacology; D000818:Animals; D018053:Arsenites / Q000494:pharmacology; D001692:Biological Transport; D002412:Cations; D002417:Cattle; D004396:Coloring Agents / Q000494:pharmacology; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005690:Galactose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D016978:Mannheimia haemolytica / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000493:pharmacokinetics*; D012709:Serum Albumin / Q000378:metabolism; D017670:Sodium Compounds / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors; D014475:Uncoupling Agents / Q000494:pharmacology
D018053:Arsenites; D002412:Cations; D004396:Coloring Agents; D012709:Serum Albumin; D017670:Sodium Compounds; D014475:Uncoupling Agents; C017947:sodium arsenite; D004024:Dicyclohexylcarbodiimide; D019158:N-Acetylneuraminic Acid; D005947:Glucose; D019297:2,4-Dinitrophenol; D005690:Galactose