Purification of the extracellular acidic proteases of Dichelobacter nodosus.
Dichelobacter nodosus, a Gram negative obligate anaerobe and causative organism of ovine footrot, secretes a family of extracellular acidic serine proteases with pI's in the range of 5.2 to 5.6, and a basic serine protease with a pI of approximately 9.5. Four acidic proteases (V1, V2, V3 and V5) from virulent and five acidic proteases (B1 to B5) from benign strains of D. nodosus were purified by chromatography on Sephadex G-100 and DEAE-Sepharose CL-6B. Proteases V2, V5 and B5 were found to yield two forms (a and b) on purification which probably arise from limited autolysis of the parent molecule. Amino acid compositions, peptide profiles produced on autolysis and apparent Mr on SDS-PAGE of proteases V1-V3 showed that they were similar to each other and to proteases B1 to B4, and that these proteases were clearly distinct from proteases V5 and B5, which were found to be identical.
Kortt
A A
AA
CSIRO, Division of Biomolecular Engineering, Parkville, Victoria, Australia.
Burns
J E
JE
Vaughan
J A
JA
Stewart
D J
DJ
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001439:Bacteroides / Q000201:enzymology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005110:Extracellular Space / Q000737:chemistry / Q000201:enzymology; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology*; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*
D000596:Amino Acids; D007527:Isoenzymes; D012697:Serine Endopeptidases; C082170:acidic protease V5, Dichelobacter