Secreted proteases from Actinobacillus pleuropneumoniae serotype 1 degrade porcine gelatin, hemoglobin and immunoglobulin A.
It was found that 48 hour cultures of Actinobacillus pleuropneumoniae secreted proteases into the medium. Electrophoresis in polyacrylamide gels (10%) copolymerized with porcine gelatin (0.1%), of the 70% (NH4)2SO4 precipitate from the culture supernatants, displayed protease activities of different molecular weights: > 200, 200, 90, 80, 70 and 50 kDa. They had activity over a broad range of pHs (4-8), with an optimal pH of 6-7. All were inhibited by 10 mM EDTA, and reactivated by 10 mM calcium. They were stable at -20 degrees C for more than a month. The proteases also degraded porcine IgA and porcine, human, and bovine hemoglobin, although they appeared to be less active against the hemoglobins. The IgA was totally cleaved in 48 h, using supernatants concentrated with polyvinyl pyrrolidone or the 70% (NH4)2SO4. Extracellular proteases could play a role in virulence.
Negrete-Abascal
E
E
Departamento de Biología Celular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico.
Tenorio
V R
VR
Serrano
J J
JJ
Garcia
C
C
de la Garza
M
M
D000189:Actinobacillus Infections / Q000378:metabolism / Q000662:veterinary; D016977:Actinobacillus pleuropneumoniae / Q000201:enzymology*; D000818:Animals; D003126:Colostrum / Q000276:immunology; D003470:Culture Media; D010450:Endopeptidases / Q000378:metabolism*; D005780:Gelatin / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007070:Immunoglobulin A / Q000378:metabolism*; D007071:Immunoglobulin A, Secretory / Q000378:metabolism; D008169:Lung Abscess / Q000382:microbiology / Q000662:veterinary; D011480:Protease Inhibitors / Q000494:pharmacology; D015203:Reproducibility of Results; D013379:Substrate Specificity; D013552:Swine; D013553:Swine Diseases / Q000378:metabolism
D003470:Culture Media; D006454:Hemoglobins; D007070:Immunoglobulin A; D007071:Immunoglobulin A, Secretory; D011480:Protease Inhibitors; D005780:Gelatin; D010450:Endopeptidases